Business, development and inclusive supply chains. Exploring pathways for the sustainable incorporation of the poor as suppliers in agrifood supply chains

El panorama global está cambiando, y esto influye sobre la forma en la que entendemos y tratamos de alcanzar un desarrollo humano sostenible. El crecimiento de la población conlleva una mayor presión sobre los recursos, pero también supone una mayor cantidad de mano de obra y de talento; la concentración en áreas urbanas está cambiando las dinámicas sociales y desafiando los canales de comercialización tradicionales, pero también genera nuevos mercados y fomenta la innovación; los cambios en la economía global están reduciendo los tradicionales desequilibrios de poder entre los países occidentales y el resto del mundo; y las crecientes interconexiones crean nuevos riesgos pero también oportunidades para lanzar iniciativas de alcance global. Todas estas tendencias nos están obligando a repensar qué es el desarrollo humano y de qué manera deberíamos afrontar el reto de la pobreza. Es comúnmente aceptado que la globalización implica interdependencia y que, para conseguir un desarrollo humano sostenible, la colaboración entre actores de distintos ámbitos es necesaria. Se observa una creciente convergencia de temas, intereses y soluciones en torno al desarrollo sostenible, incluso en diferentes países y sectores, lo que está facilitando la colaboración estratégica entre empresas, gobiernos y sociedad civil. Existen pocas duda a día de hoy sobre el papel fundamental que las empresas deben desempeñar en la transición mundial hacia la sostenibilidad ambiental y la erradicación de la pobreza. Las empresas están evolucionando desde un enfoque tradicional centrado en la maximización de beneficios económicos hacia un enfoque holístico que integra la sostenibilidad y la responsabilidad social como parte del núcleo de negocio de las compañías. En el ámbito medioambiental, muchas empresas ya han comenzado a actuar y tratan de reducir sus emisiones, sus desechos y su consumo de energía. Sin embargo la contribución de las empresas a la reducción de la pobreza no está tan clara. Actualmente en torno a 1,2 miles de millones de personas viven en situación de extrema pobreza. La mayoría de estas personas aún vive en zonas rurales donde la mayor parte de la población activa trabaja en el sector agrícola. Por lo tanto, mejorar las oportunidades y reducir los riesgos de los productores más vulnerables en el sector de la agricultura puede ser un motor de desarrollo rural y reducción de la pobreza, especialmente en países de bajo nivel de desarrollo cuyas economías están fundamentalmente basadas en la agricultura. Algunas empresas comienzan a incluir a los pobres en sus operaciones como consumidores, proveedores y emprendedores. Esta tesis se centra en las potenciales oportunidades relacionadas con la incorporación sostenible de los pobres como proveedores de productos y/o de mano de obra. La colaboración entre empresas y productores vulnerables de países en desarrollo es un tema relativamente nuevo y todavía poco estudiado. La pregunta que guía esta tesis es: “¿Cómo pueden las empresas facilitar la inclusión sostenible en cadenas de suministro de productores vulnerables de los países menos desarrollados?”. Para responder a la pregunta anterior, la autora ha aplicado una metodología de casos de estudio. Esta metodología se considera apropiada porque la investigación sobre cadenas de suministro inclusivas es todavía escasa y porque es necesario entender en profundidad un fenómeno de la vida real, y para ello es fundamental conocer su contexto. En primer lugar, se realiza una revisión de literatura para identificar las proposiciones y los constructos teóricos que guiarán la posterior recogida de datos. La revisión de literatura se divide en dos partes: una más general que explora la dimensión social de la sostenibilidad en cadenas de suministro, y una más específica que se centra en la incorporación de los pobres como proveedores en cadenas de suministro. A lo largo de la última década, ha habido un crecimiento exponencial de los estudios académicos sobre la sostenibilidad de las cadenas de suministro, pero la mayoría de los esfuerzos se han dirigido hacia la dimensión medioambiental de la sostenibilidad. Por lo tanto la revisión de literatura, que se presenta en la Sección 3.1 (página 35) y que profundiza en la sostenibilidad social de las cadenas de suministro, puede considerarse una contribución en sí misma. Esta revisión de literatura revela que la investigación sobre aspectos sociales en cadenas de suministro está cobrando impulso en distintas áreas de conocimiento, principalmente en los ámbitos de investigación sobre “gestión de cadenas de suministro”, “responsabilidad social corporativa” y “estudios del desarrollo”. La investigación existente sobre sostenibilidad social de cadenas de suministro se centra en tres temas: aclarar la definición de sostenibilidad social; analizar la implementación de estrategias de sostenibilidad social en cadenas de suministro; y estudiar el apoyo de las em presas líderes a proveedores vulnerables para facilitar su transición hacia la sostenibilidad. Un marco conceptual que resume los principales hallazgos de esta primera parte de la revisión de literatura es planteado en la Figura 7 (página 48). No obstante, en el área de investigación que está emergiendo en torno a la sostenibilidad social de las cadenas de suministro, los estudios relacionados con la reducción de la pobreza son aún escasos. Además se aprecia una falta de contribuciones desde y sobre los países menos desarrollados, así como una clara tendencia a reflejar la visión de las empresas líderes de las cadenas de suministro, olvidando la perspectiva de los proveedores. La segunda parte de la revisión de literatura presentada en la Sección 3.2 (página 51) profundiza en tres líneas de investigación que exploran, desde distintas perspectivas, la inclusión de los pobres en cadenas de suministro. Estas líneas son “Global Value Chains” (GVC), “Base of the Pyramid” (BoP) y “Sustainable Supply Chain Management” (SSCM). La investigación en GVC analiza las cadenas de suministro desde la perspectiva de la globalización económica y el comercio internacional, poniendo especial énfasis en las implicaciones para los países en desarrollo y las comunidades vulnerables. GVC caracteriza las cadenas de suministro según la forma en la que son gobernadas, las oportunidades de mejora que existen para los productores que forman parte de la cadena y el grado de inclusión o exclusión de las comunidades más pobres y vulnerables. La investigación en BoP explora las relaciones comerciales entre empresas y comunidades pobres. La premisa fundamental del concepto BoP es la posibilidad de combinar la generación de beneficios con la reducción de la pobreza. La propuesta original es que mediante la venta de productos y servicios a las comunidades pobres de países en desarrollo, la pobreza puede ser reducida al tiempo que las empresas incrementan sus beneficios, especialmente las grandes empresas multinacionales. Esta idea ha ido evolucionando y, a día de hoy, los investigadores BoP consideran la incorporación de los pobres no sólo como consumidores sino también como empleados, proveedores y co-creadores. La investigación en SSCM ha estado fundamentalmente orientada al estudio de la dimensión medioambiental de la sostenibilidad de cadenas de suministro. Sin embargo, la creciente externalización de la producción a países en desarrollo y las demandas de los grupos de interés para que las empresas aborden todos los aspectos de la sostenibilidad han llevado a los académicos de SSCM a reconocer la importancia de integrar asuntos relacionados con la reducción de la pobreza en sus investigaciones. Algunos estudios comienzan a apuntar los principales retos a los que se enfrentan las empresas para colaborar con productores vulnerables en sus cadenas de suministro. Estos retos son: falta de comunicación, altos costes de transacción y el incremento de la complejidad de las operaciones. Las contribuciones de estas tres líneas de investigación son complementarias para el estudio de las cadenas de suministro inclusivas. Sin embargo, raramente han sido consideradas conjuntamente, ya que pertenecen a ámbitos de conocimiento distintos. Esta tesis integra las aportaciones de GVC, BoP y SSCM en un marco conceptual para la creación y gestión de cadenas de suministro inclusivas. Este marco conceptual para cadenas de suministro inclusivas queda representado en la Figura 9 (página 68). El marco conceptual refleja las motivaciones que llevan a las empresas a colaborar con productores vulnerables, los retos a los que se enfrentan al hacerlo, y los caminos o estrategias que están siguiendo para construir y operar cadenas de suministro inclusivas de manera que sean beneficiosas tanto para la empresa como para los productores vulnerables. A fin de validar y refinar el marco conceptual propuesto, tres casos de estudio se llevan a cabo. Las cadenas de suministro analizadas por los casos de estudio pertenecen al sector agrícola y sus principales proveedores se encuentran en países de África subsahariana. Múltiples métodos de recolección de datos y triangulación son utilizados para mejorar la fiabilidad de los datos. La autora desarrolló trabajos de campo en Senegal, Etiopía y Tanzania. Estos viajes permitieron enriquecer el proceso de recogida de información mediante entrevistas semiestructuradas y conversaciones informales con los principales actores de la cadena de suministro y mediante la observación directa de los procesos y las interacciones entre productores vulnerables y empresas. El Caso de estudio A (Sección 5.1 en página 96) es un caso de estudio único. Analiza la cadena de suministro local de verduras en Senegal. La organización focal es Manobi, una empresa senegalesa que provee servicios de bajo coste a emprendedores locales del sector agrícola. El Caso de estudio A proporciona un interesante análisis del funcionamiento de una cadena de suministro local en un país en desarrollo y muestra como la provisión de servicios profesionales puede mejorar el desempeño de productores vulnerables. El Caso de estudio B (Sección 5.2 en página 122) es un caso de estudio único. Analiza la cadena de suministro global de flor cortada con origen en Etiopía. La organización focal es EHPEA, la Asociación Etíope de Productores y Exportadores Hortícolas, cuya misión es promover y salvaguardar la posición competitiva del sector agrícola etíope en el mercado global. El Caso de estudio B ayuda a comprender mejor la perspectiva de los proveedores respecto a los requerimiento de sostenibilidad del mercado global. También muestra cómo la inclusión de los productores en el proceso de desarrollo de un estándar privado facilita su implementación posterior. El Caso de estudio C (Sección 5.3 en página 143) es un caso de estudio múltiple. Analiza la cadena de suministro global de café especial con origen en Tanzania. Las organizaciones focales son comerciantes que conectan de manera directa a pequeños agricultores de café en países en desarrollo con empresas tostadoras de café en países desarrollados. El Caso de estudio C muestra cómo un pequeño agricultor puede proveer un producto “premium” al mercado global, y participar en un segmento diferenciado del mercado a través de una cadena de suministro transparente y eficiente. Las aportaciones empíricas de los casos de estudio ayudan a validar y mejorar el marco conceptual sobre cadenas de suministro inclusivas (ver discusión en el Capítulo 6 en página 170). El resultado es la propuesta de una nueva versión del marco conceptual representado en la Figura 40 (página 195). Los casos de estudio también proporcionan interesantes aportaciones en relación a la gestión de cadenas de suministro inclusivas y muestran las perspectivas de distintos actores implicados. Esta tesis arroja luz sobre el papel de las empresas en la creación y la gestión de cadenas de suministro inclusivas llevando a cabo una revisión de literatura multidisciplinar y analizando tres casos de estudio en países africanos. Como resultado, esta tesis presenta una serie de contribuciones empíricas y teóricas al ámbito de investigación emergente en torno a las cadenas de suministro inclusivas (Capítulo 7). Esta tesis también pretende ser útil a profesionales que deseen facilitar la incorporación de los pobres como proveedores en condiciones justas y beneficiosas. ABSTRACT The global outlook is changing, and this is influencing the way we understand and try to achieve sustainable human development. Population growth entails increasing pressure over resources, but it also provides greater workforce and talent; concentration in urban areas is changing social dynamics and challenging traditional marketing channels, but also creating news markets and driving innovation; the global economy shift is rebalancing the traditional power imbalance between Western countries and the rest of the world, making new opportunities to arise; and interconnections and global interdependence create new risks but also opportunities for launching initiatives with a global reach. All these trends are impelling us to rethink what development is and in which way poverty alleviation should be approached. It is generally agreed that globalization implies interdependence and, in order to achieve sustainable human development, collaboration of all actors is needed. A convergence of issues, interests and solutions related to sustainable development is being observed across countries and sectors, encouraging strategic collaboration among companies, governments and civil society. There is little doubt nowadays about the crucial role of the private sector in the world’s path towards environmental sustainability and poverty alleviation. Businesses are evolving from a “business as usual” stance to a more sustainable and responsible approach. In the environmental arena, many companies have already “walk the talk”, implementing environmental management systems and trying to reduce emissions and energy consumption. However, regarding poverty alleviation, their contribution is less clear. There are around 1.2 billion people living in extreme poverty. Most of this people still live in rural areas where the agricultural sector employs a big part of the active population. Therefore, improving opportunities and reducing risks for vulnerable producers in the agri-food sector can be a primary engine of rural development and poverty alleviation, particularly in the poor, agriculture-based economies of least developed countries. Some companies are beginning to include the poor into their operations as consumers, suppliers and entrepreneurs. This thesis focuses specifically on the potential opportunities related to the sustainable incorporation of the poor as suppliers of products and/or labor. Business collaboration with vulnerable producers in developing countries is a relatively new trend and it is still understudied. The overall question guiding this thesis is: “How can businesses facilitate the sustainable inclusion of vulnerable producers from least developed countries into supply chains?”. In order to answer the research question, the author has applied a case study research strategy. This methodology is considered appropriate because research about inclusive supply chains is still at an early stage, and because there is a need to understand a real-life phenomenon in depth, but such understanding encompasses important contextual conditions. First, a literature review is conducted, in order to identify the research propositions and theoretical constructs that will guide the data collection. The literature review is divided in two parts: a more general one that explores the social dimension of sustainability of supply chains, and a more specific one that focuses on the incorporation of the poor as suppliers in supply chains. During the last decade, there has been an exponential growth of studies in the field of supply chain sustainability, but research efforts have traditionally been directed towards the analysis of the environmental dimension. Therefore, the literature review presented in Section 3.1 (page 35) that delves into social sustainability of supply chains can be considered a contribution in itself. This literature review reveals that the investigation of social issues in supply chains is gaining momentum and comes from different academic disciplines, namely Supply Chain Management, Corporate Social Responsibility and Development Studies. Existing research about social sustainability of supply chains focuses on three issues: clarify the definition of social sustainability; analyze the implementation of social sustainability strategies in supply chains; and study lead companies’ support to vulnerable suppliers in their transition towards sustainability. A conceptual framework that outlines the main findings that emerge from this first part of literature review is proposed in Figure 7 (page 48). Nevertheless, in this nascent field of social sustainability of supply chains, studies related to poverty alleviation are still scarce. Moreover, a lack of contributions from and about least developed countries has been observed, as well as a tendency to reflect on the lead firms’ standpoint, neglecting the suppliers’ perspective. The second part of the literature review (Section 3.2 in page 51) delves into three research streams that are exploring the inclusion of the poor into supply chains from different viewpoints. These research streams are Global Value Chains (GVC), Base of the Pyramid (BoP) and Sustainable Supply Chain Management (SSCM). GVC research discusses the dynamics of economic globalization and international trade, putting special emphasis in the implications for developing countries and vulnerable communities. GVC characterizes supply chains by the way they are governed, the upgrading opportunities that exist for producers in the chain and the degree of inclusion or exclusion of impoverished communities. BoP research explores trading relationships between businesses and impoverished communities. The core premise of the BoP concept is the possibility to combine profits with poverty alleviation. The original BoP proposition is that by marketing innovative products and services to poor communities in developing countries, poverty would be reduced and companies would increase their benefits, especially multinational companies. This idea has evolved to consider the incorporation of the poor to business activities not only as consumers, but also as employees, entrepreneurs and co-creators. The SSCM school of thought has mainly focused on studying the environmental dimension of supply chain sustainability, neglecting the consideration of the social perspective. However, in recent years, increasing outsourcing of production to developing countries and stakeholders’ demands for a more holistic approach to business sustainability have led SSCM scholars to acknowledge the importance of integrating poverty concerns in this field’s research agenda. Some SSCM studies identify the main operational challenges for companies which engage with vulnerable suppliers in their supply chains: missing communication, higher transactional and operational costs and increased complexity. Contributions from these three research streams are complementary for the study of inclusive supply chains. However, they have been rarely considered together, since they belong to different research areas. This thesis seeks to play a dovetailing role in this scenario by proposing a conceptual framework for creating and operating inclusive supply chains that builds on contributions from GVC, SSCM and BoP research. This framework for inclusive supply chains is depicted in Figure 9 (page 68), and explains the motivations that drive businesses to collaborate with vulnerable suppliers, the chal lenges they face in doing so, and the pathways they are following in order to build and operate inclusive supply chains profitably for both buying companies and vulnerable suppliers. In order to validate and refine the proposed framework, three case studies are carried out. The supply chains analyzed by the case studies belong to the agri-food sector and source from Sub-Saharan African countries. Multiple data collection methods and triangulation are used in order to improve reliability of findings. The author carried out field work in Senegal, Ethiopia and Tanzania. These travels enriched the data collection process, providing semi-structured interviews and informal conversations with the main actors in the supply chains, as well as direct observation of processes and interactions among companies and vulnerable suppliers. Case study A (Section 5.1 in page 96) is a single case study. It analyzes a local supply chain in Senegal providing vegetables to the local market. The focal organization is Manobi, a Senegalese inclusive business which provides affordable ICT services to local entrepreneurs in the agri-food sector. Case study A provides interesting insights into the dynamics of local supply chains and how professional services can help to improve their performance. Case study B (Section 5.2 in page 122) is a single case study. It analyzes a global supply chain with origin in Ethiopia providing cut flowers to the global commodity market. The focal organization is EHPEA, Ethiopian Horticulture Producers and Exporters Association, whose mission is to promote and safeguard the competitive position of the Ethiopian horticulture sector within the global market. Case study B helps to better understand the suppliers’ perspective regarding global market sustainability requirements and shows how the inclusion of suppliers in the process of development of a private standard has a positive impact in its implementation. Case study C (Section 5.3 in page 143) is a multiple case study. It analyzes a global supply chain with origin in Tanzania providing coffee to the global niche market of specialty coffee. The focal organizations are traders who are directly connecting smallholder coffee farmers in developing countries to coffee roasters in developed countries. Case study C shows how smallholder farmers can supply a premium product and be incorporated in a differentiated market segment through a transparent and efficient supply chain. The empirical findings from the case studies help to validate and refine the conceptual framework (see discussion in Chapter 6). The proposal of a new version of the conceptual framework is depicted in Figure 40 (page 195). The case studies also provide interesting insights related to the management of inclusive supply chains and show the perspectives of the different actors involved. This thesis sheds some light on the role of businesses in the creation and operation of inclusive supply chains by carrying out a cross-disciplinary literature review and analyzing three case studies in African countries. In doing so, this thesis presents a series of theoretical and empirical contributions to the emerging academic field of inclusive supply chains (Chapter 7). This thesis also intends to be useful to practitioners willing to improve the incorporation of the poor as suppliers in fair and profitable conditions.

Rem : a los dos lados del espejo = (Rem at both sides of the mirror)

Cuando se trata de Rem Koolhaas, su espejo no refleja una sola imagen sino múltiples, es un prisma poliédrico. Su espejo nos devuelve el Rem mediático, el intelectual, el conceptualizador, el constructor, el analista, el periodista, el actor... En el caso de esta investigación, fijamos el punto de mira en el Rem COMUNICADOR. “Rem a los dos lados del espejo” se enmarca en una investigación sobre los medios de comunicación de arquitectura, su reflejo en la producción arquitectónica y viceversa. Se trata de llegar a discernir si comunicación y producción arquitectónica colisionan y confluyen en el caso de grandes comunicadores como Rem Koolhaas, si el mensaje y el medio transmisor adquieren las mismas cualidades. Centrándose en la figura de Rem Koolhaas, la tesis aborda la evolución de su faceta comunicativa y las transformaciones sucesivas en el campo de la comunicación arquitectónica, en paralelo a su evolución conceptual a lo largo de su trayectoria. La investigación, por tanto, no se centra tanto en su componente teórica o en la práctica arquitectónica de OMA, sino en la exposición de su producción al mundo, especialmente a través de sus ensayos y libros. “Delirious New York” y “SMLXL” son un reflejo del momento conceptual en que se inscriben, y contienen mucha información sobre los referentes gráficos que irremediablemente han influido en su composición. Especialmente, la aparición de “SMLXL” supuso un revulsivo para el mundo de la comunicación arquitectónica, porque puso el foco sobre la importancia de dejar atrás un discurso narrativo linea y unifocal, para afrontar la comunicación barajando múltiples variables, y aproximaciones, en un proceso similar al desarrollo de un proyecto de arquitectura. Presenta un diseño muy novedoso y una edición extremadamente cuidada, que atiende a parámetros mucho más ambiciosos que los meramente narrativos. Profundiza en la necesidad de una temática global, planteando cuál es la aproximación más apropiada para cada uno de los proyectos que describe, transmitiendo al lector una percepción más allá de lo estrictamente visual, más próximo a lo sensorial. Además, su enorme repercusión a nivel internacional y el gran interés que despertó (no solamente entre los arquitectos, sino también entre diseñadores gráficos, publicistas, personas provenientes de todo tipo de tendencias artísticas y público en general), provocó la globalización del fenómeno de las publicaciones arquitectónicas y puso de manifiesto la importancia de la comunicación como una disciplina en sí misma, dentro de la producción arquitectónica en la era actual. A pesar de la importancia de “SMLXL” a todos los niveles, la presente tesis plantea que, donde realmente se culmina esa experiencia comunicativa, es en “Content”, al incluir nuevos parámetros relacionados con la fusión conceptual de continente y contenido. Es en esta publicación donde el objeto de la comunicación y la expresión de la misma se convierten en un único elemento, que se rige por leyes similares. En este caso, la ley fundamental es la aplicación hasta sus máximas consecuencias de la “cultura de la congestión”, tanto en el mensaje como en el medio, generando lo que hemos convenido en denominar “comunicación congestiva”. Esta concepción deviene en que necesariamente se materialice como un producto efímero, desechable, casi virtual, porque responde a las condiciones de un momento muy concreto y específico y fuera de ese contexto pierde su significación.. La “cultura de la congestión” empieza a surgir en los planteamientos de Koolhaas en la Architectural Association School of Architecture de Londres, bajo la tutela de Elia Zenghelis. Posteriormente se desarrolla en su manifiesto retroactivo sobre Manhattan, “Delirious New York”, donde declara la guerra abierta al urbanismo del movimiento moderno y afirma que la ciudad realmente contemporánea es aquella que es fruto de un desarrollo no planificado, hiperdensa y posible gracias a los avances tecnológicos de su era. Finalmente comienza a materializarse en la Diploma Unit 9 de la AA, donde entra como profesor en 1975, dejando una huella indeleble en las generaciones posteriores de arquitectos que pasaron dicha unidad. Rem Koolhaas es ante todo un intelectual y por ello, todo el constructo teórico sobre la metrópolis comienza a reflejarse en su obra a través de OMA desde el comienzo de su producción. Podemos decir a grandes rasgos que su carrera está marcada por dos hitos históricos fundamentales que determinan tres etapas diferenciadas en su producción. En sus primeros años de profesión, Koolhaas sigue fascinado por la metrópolis urbana y la aplicación del método paranoico crítico a su producción arquitectónica. Es un arquitecto profundamente surrealista. Entiende este método como una estrategia de conocimiento y aproximación al mundo que le rodea: “dejar salir el inconsciente pero sostenerlo con las muletas de la racionalidad”. Pero lo que en realidad le interesa es su aplicación a la gran escala, el “Bigness”, y por ello, participa en proyectos muy ambiciosos de los que surgen conceptos que, más allá de resultar premiados o no, han dejado una huella ideológica en el devenir de la arquitectura. Entre estos proyectos, cabe destacar su propuesta para el Parque de la Villette o la Très Grande Bibliotèque de París. Sus proyectos de esta época destilan una gran carga conceptual, que devienen en unos interiores sorprendentes pero una apariencia exterior sobria o incluso podríamos decir "povera", por el uso de materiales efímeros, poco habituales en la macro-arquitectura hasta ese momento. Súbitamente, en 1997, explotó el denominado “Efecto Bilbao”, de la mano de Frank Gehry (1). El Museo Guggenheim de Bilbao, con su espectacularidad, sus formas pregnantes e imposibles, impacta al mundo. Nace la era de la “Arquitectura del Espectáculo”; la transformación de la ciudad a través de ICONOS que actúen como nodos de atracción y concentración en torno a los cuales supuestamente se revitaliza la actividad económica, cultural y sociopolítica de la ciudad, como si a través de un único gesto se pudieran regenerar todos los tejidos internos de la urbe. Rem Koolhaas comprende rápidamente que la aproximación a la ciudad ha cambiado y, sobre todo, el mercado. En el mundo de la globalización, la única manera de llegar a materializar el “Bigness”, es encerrando sus ejercicios intelectuales en formas pregnantes, bellas, icónicas, espectaculares. Koolhaas encuentra su marca personal en la estética “Stealth”, proveniente de los aviones de combate facetados para evitar los radares, elaborados en los años 80. De esta época surgen proyectos como la Casa da Música de Oporto o la Biblioteca de Seattle; ambos edificios son iconos facetados, de belleza pregnante, que dejan una huella indeleble en la ciudad y provocan, al igual que el Guggenheim, un cierto efecto de recuperación y revitalización en el entorno en que se asientan, al menos de manera temporal. En cualquier caso, Koolhaas nunca abandona los ejercicios meramente teóricos, pero segrega su actividad en dos: OMA produce aquello que tiene vocación de ser construido y se rige por los parámetros del mercado global y AMO, la otra cara del espejo de Rem, aplica el pensamiento arquitectónico a campos no explorados, sin la dependencia de agentes externos, pudiendo permitirse ser un laboratorio puramente experimental. En este escenario, llega el 11 de septiembre de 2001 y el ataque a las Torres Gemelas de Nueva York tiene efectos devastadores a todos los niveles, significando, en un período de tiempo sorprendentemente corto, un cambio en el orden mundial. Rem Koolhaas da entonces un giro de 180 grados, dirige su mirada hacia China, donde entiende que sus aportaciones tienen un beneficio social más directo que en occidente. (2) Para presentar al mundo su nuevo cambio de rumbo y la creación del “Think Tank” AMO, plantea una gran exposición en la NeueGallerie de Berlín bajo el título de “Content”, experiencia paralela a la edición del libro con el mismo título, que inicialmente nace como “catálogo de la exposición, pero que internamente siempre se concibió como el documento más trascendente en el estudio desde “SMLXL”. Sin embargo, en muchos aspectos se trata de su opuesto: una publicación con formato revista, de tapa blanda, con paginado muy fino, formato de "folleto de supermercado" y contenido hiperdenso. Es un experimento efímero, fugaz, ligero, barato, de “usar y tirar”. De hecho, está fuera de stock, ya no se edita. Probablemente Rem Koolhaas desaprobaría que se hiciera una investigación que pusiera el foco sobre el mismo, porque diez años después de su publicación seguramente opine que su vigencia ha caducado. Sin embargo, muestra con una claridad meridiana el estado conceptual y vital de OMA en el momento de su publicación y representa, además un verdadero hito en la comunicación arquitectónica, un punto de no retorno, el máximo exponente de lo que hemos denominado “comunicación congestiva”. La presente tesis plantea que “Content” contiene la esencia de la mayor aportación de Rem Koolhaas al mundo de la arquitectura: la transformación profunda y definitiva de la comunicación arquitectónica mediante la convergencia del estado conceptual y la transmisión del mismo. Su legado arquitectónico y conceptual ha marcado a todas las generaciones posteriores de manera indeleble. Sus ensayos, sus teorías, sus proyectos y sus edificaciones ya pertenecen a la historia de la arquitectura, sin ninguna duda. Pero es su revisión del concepto de la comunicación en arquitectura lo que ha tenido y tendrá un reflejo inmediato en las generaciones futuras, no solamente en la comunicación sino en su arquitectura, a través de un intercambio biyectivo. El planteamiento a futuro sería determinar qué sucede tras “Content”, tras la hiperdensidad máxima, tras la cultura de la congestión visual; qué es lo que propone Koolhaas y qué se va a plantear también en el mundo de la comunicación arquitectónica. Para ello, estudiaremos en profundidad sus últimos proyectos relacionados con la comunicación, como su propuesta para la Biennale de Arquitectura de Venecia de 2014, su intensa investigación sobre el “Metabolismo” en “Project Japan: Metabolism Talks...”, o la dirección de sus últimos planteamientos territoriales. En los últimos tiempos Rem Koolhaas habla de “Preservación”, de “Sobriedad”, de “Esencialismo”, de “Performance”... El autor intelectual de la cultura de la congestión habla ahora de la “low density”...como no podía ser de otra manera en la otra cara del espejo. En definitiva, el color blanco como suma de todos los colores, todas las longitudes de onda del espectro visible recibidas al tiempo. ABSTRACT When talking about Rem Koolhaas, the mirror does not only reflect one but numerous images: it is nothing but a polyhedral prism. His mirror gives us the image of Rem the media celebrity, the intellectual, the conceptualizer, the builder, the analyst, the journalist, the actor... This research sets the spotlight on Rem the COMMUNICATOR. "Rem on both sides of the mirror" belongs to a research on architectural media, its influence on the architectural production and vice versa. It is aimed at getting to discern whether communication and architectural production collide and converge in the case of great communicators such as Rem Koolhaas, and whether the message and transmission media acquire the same features. Focusing on the figure of Rem Koolhaas, this thesis addresses the evolution of his communicative facet and the successive transformations in the field of architectural communication, parallel to the conceptual evolution he underwent throughout his career. Therefore, this research is not so much focused on his theoretical component or on the OMA’s architectural practice, but on the exhibition of his production to the world, especially through his essays and books. "Delirious New York" and "SMLXL" hold up a mirror to the conceptual moment they are part of, and contain a great deal of information about the graphic references that have inevitably influenced his work. Specially, the launch of "SMLXL" was a salutary shock for the architectural communication world, since it set the spotlight on the importance of leaving a linear and unifocal narrative behind in order to face communication considering multiple variables and approaches, based on a process similar to the development of an architectural project. It offers a very innovative design and an extremely careful editing, which deals with parameters much more ambitious than those merely narrative. It explores the need for a global subject and suggests the most appropriate approach for each of the projects described, giving the reader a closer insight to the sensory that goes beyond what’s strictly visual. In addition, its huge international impact and the great interest shown, not only by architects but also by graphic designers, publishers, people from all kinds of artistic trends and the general public, led to the globalisation of the architectural publications phenomenon and brought the importance of communication as a discipline in itself, within the architectural production in the age at hand, to light. Despite the importance of "SMLXL" at all levels, this thesis suggests that the communication experience really culminates in "Content", for it includes new conceptual parameters associated with the container-content conceptual fusion. It is in this book where the purpose of communication and the expression of such become a single element, ruled by similar laws. In this particular case, the fundamental law is to implement the "culture of congestion" to its extreme consequences in both the message and the media, leading to what we have agreed to refer to as "congestive communication”. This concept leads to its inevitable materialisation into an ephemeral, disposable, almost virtual product, because it meets the conditions of a very concrete and specific time, and outside that context it loses its significance. The "culture of congestion" emerged in Koolhaas’ approaches under the guidance of Elia Zenghelis, in the Architectural Association School of Architecture of London. Subsequently, his retroactive manifesto on Manhattan, "Delirious New York" developed it, waging an all-out war against the modern movement urbanism and maintaining that the really contemporary cities are those hyperdense ones that rise as a result of an unplanned development and thanks to the typical technological advances of their time. Finally it began to materialise in the Diploma Unit 9 of the AA, in which he started lecturing in 1975, leaving an indelible mark on subsequent generations of architects who passed that unit. First and foremost, Rem Koolhaas is an intellectual and, therefore, all the theoretical construct in the metropolis began to be reflected in his work through OMA since the beginnings of his production. Broadly speaking, we can say that his career is influenced by two essential historic events, which determine three different stages in his production. In the early years of his career, Koolhaas was still fascinated by the urban metropolis and the implementation of the paranoiac-critical method to his architectural production. He was then a deeply surreal architect. He understood this method as a knowledge strategy and an approach to the world around him: "let the subconscious out but hold it with the crutches of reasonableness”. However, he was actually interested in its implementation on a broad scale, the "Bigness", and therefore, he took part in ambitious projects that led to the accrual of concepts that, beyond being rewarded, left an ideological impression on the evolution of architecture. These projects included his proposal for the Parc de la Villette or the Très Grande Bibliotèque in Paris. The projects he carried out during this period showed a great conceptual background, which evolved into surprising interiors but a sober, or even "povera", exterior appearance, thanks to the use of ephemeral materials that were atypical in the macro-architecture field until that moment. Suddenly, in 1997, the so-called "Bilbao effect" boomed thanks to Frank Gehry (1). The Guggenheim Museum of Bilbao amazed the world with its spectacular nature and its pregnant and impossible shapes. It was the beginning of the era of “The architecture of spectacle”: the transformation of the city through ICONS that would act as nodes of attraction and gathering, around which the economic, cultural and socio-political activity of the city was supposed to be revitalized, as if through a single gesture all internal tissues of the city could be rebuilt. Rem Koolhaas quickly realized that the approach to the city, and especially to the global market, had changed. In the world of globalisation, the only way to get to materialise such "Bigness" was by keeping his intellectual exercises in pregnant, beautiful, iconic and spectacular shapes. Koolhaas found his personal brand in the Stealth aesthetic, resulting from the eighties American combat aircrafts whose shape was faceted in order to avoid radars. Projects such as the Casa da Música in Porto or the Seattle Library date from this period; both buildings are faceted icons of pregnant beauty that left an indelible mark on the city and caused, like the Guggenheim, some degree of recovery and revitalization on the environment in which they were based, at least temporarily. In any case, Koolhaas never gave the merely theoretical exercises up, but he segregated his work in two: OMA produced what was destined to be built and ruled by the parameters of the global market and AMO, Rem’s other side of the mirror, applied the architectural thought in unexplored fields, notwithstanding external agents and being able to work as a purely experimental laboratory. In light of this backdrop, September 11th 2001 came and the attacks on the Twin Towers in New York had devastating effects at all levels, leading to a change in the world order, in a surprisingly short period of time. Rem Koolhaas made a 180° turn directing his vision towards China, where he believed his contributions would have a more direct social benefit than in the Western world. (2) In order to introduce his new course of direction and the creation of the AMO "Think Tank", he planned a major exhibition in the Neue Nationalgalerie of Berlin under the title "Content", in parallel with edition of the book with the same title, which was at first the "exhibition catalog” but, deep down, was always conceived as the most important document of the Office since "SMLXL". However, in many ways it was just the opposite: a publication characterised by its magazine format, soft cover, very fine paging, "supermarket brochure" form and hyperdense content. It was an ephemeral, brief, light, cheap and "disposable" experiment. In fact, it is currently out of stock and out of print. Rem Koolhaas would probably disapprove of a research that sets the spotlight on him, for he would probably say that his validity has expired given that it has been ten years since its publication. However, it shows OMA’s conceptual and vital status at the time of its publication with crystalline clarity and it is also a true milestone in architectural communication. A point of no return. The epitome of the so-called "congestive communication ". This thesis suggests that "Content" contains the essence of Rem Koolhaas’ greatest contribution to the world of architecture: the deep and definitive transformation of architectural communication through the convergence of the conceptual state and the transmission thereof. His architectural and conceptual legacy has left an indelible mark on all subsequent generations. There is no doubt his essays, theories, projects and buildings already belong to the history of architecture. But it is his review on the concept of communication in architecture that has had and shall have an immediate influence on future generations, not only in their communication but also in their architecture, through a bijective exchange. Future approaches should try to determine what happens after "Content", after the maximum hyperdensity, after the visual culture of congestion; what shall Koolhaas suggest as well as what shall happen in the world of architectural communication. To this end, we shall study his latest communication-related projects, such as the design of the Venetian Architecture Biennale in 2014, his intensive research on the "Metabolism" in "Project Japan: Metabolism Talks ...", or the course of his latest territorial approaches in depth. Most recently, Rem Koolhaas has talked about "Preservation", "Sobriety" of "Essentialism", "Performance", etc. The mastermind of the culture of congestion now speaks of the "low density"... as it could not be otherwise, on the other side of the mirror. Summarizing, the white color as the sum of all colors; all wavelengths of the visible spectrum received at the same time.

Proceso metodológico para optimizar la toma de decisiones en el proyecto de envolventes arquitectónicas multicapa = Methodological process for the optimization of the decision making in multilayer building envelope projects

La presente tesis doctoral se enmarca dentro del concepto de la sistematización del conocimiento en arquitectura, más concretamente en el campo de las construcciones arquitectónicas y la toma de decisiones en la fase de proyecto de envolventes arquitectónicas multicapa. Por tanto, el objetivo principal es el establecimiento de las bases para una toma de decisiones informadas durante el proyecto de una envolvente multicapa con el fin de colaborar en su optimización. Del mismo modo que la historia de la arquitectura está relacionada con la historia de la innovación en construcción, la construcción está sujeta a cambios como respuesta a los fracasos anteriores. En base a esto, se identifica la toma de decisiones en la fase de proyecto como el estadio inicial para establecer un punto estratégico de reflexión y de control sobre los procesos constructivos. La presente investigación, conceptualmente, define los parámetros intervinientes en el proyecto de envolventes arquitectónicas multicapa a partir de una clasificación y sistematización de todos los componentes (elementos, unidades y sistemas constructivos) utilizados en las fachadas multicapa. Dicha sistematización se materializa en una hoja matriz de datos en la que, dentro de una organización a modo de árbol, se puede acceder a la consulta de cada componente y de su caracterización. Dicha matriz permite la incorporación futura de cualquier componente o sistema nuevo que aparezca en el mercado, relacionándolo con aquellos con los que comparta ubicación, tipo de material, etc. Con base en esa matriz de datos, se diseña la sistematización de la toma de decisiones en la fase de proyecto de una envolvente arquitectónica, en concreto, en el caso de una fachada. Operativamente, el resultado se presenta como una herramienta que permite al arquitecto o proyectista reflexionar y seleccionar el sistema constructivo más adecuado, al enfrentarse con las distintas decisiones o elecciones posibles. La herramienta se basa en las elecciones iniciales tomadas por el proyectista y se estructura, a continuación y sucesivamente, en distintas aproximaciones, criterios, subcriterios y posibilidades que responden a los distintos avances en la definición del sistema constructivo. Se proponen una serie de fichas operativas de comprobación que informan sobre el estadio de decisión y de definición de proyecto alcanzados en cada caso. Asimismo, el sistema permite la conexión con otros sistemas de revisión de proyectos para fomentar la reflexión sobre la normalización de los riesgos asociados tanto al proprio sistema como a su proceso constructivo y comportamiento futuros. La herramienta proporciona un sistema de ayuda para ser utilizado en el proceso de toma de decisiones en la fase de diseño de una fachada multicapa, minimizando la arbitrariedad y ofreciendo una cualificación previa a la cuantificación que supondrá la elaboración del detalle constructivo y de su medición en las sucesivas fases del proyecto. Al mismo tiempo, la sistematización de dicha toma de decisiones en la fase del proyecto puede constituirse como un sistema de comprobación en las diferentes fases del proceso de decisión proyectual y de definición de la envolvente de un edificio. ABSTRACT The central issue of this doctoral Thesis is founded on the framework of the concept of the systematization of knowledge in architecture, in particular, in respect of the field of building construction and the decision making in the design stage of multilayer building envelope projects. Therefore, the main objective is to establish the bases for knowledgeable decision making during a multilayer building envelope design process, in order to collaborate with its optimization. Just as the history of architecture is connected to the history of innovation in construction, construction itself is subject to changes as a response to previous failures. On this basis, the decisions made during the project design phase are identified as the initial state to establish an strategic point for reflection and control, referred to the constructive processes. Conceptually, this research defines the parameters involving the multilayer building envelope projects, on the basis of a classification and systematization for all the components (elements, constructive units and constructive systems) used in multilayer façades. The mentioned systematization is materialized into a data matrix sheet in which, following a tree‐like organization, the access to every single component and its characterization is possible. The above data matrix allows the future inclusion of any new component or system that may appear in the construction market. That new component or system can be put into a relationship with another, which it shares location, type of material,… with. Based on the data matrix, the systematization of the decision making process for a building envelope design stage is designed, more particularly in the case of a façade. Putting this into practice, it is represented as a tool which allows the architect or the designer, to reflect and to select the appropriate building system when facing the different elections or the different options. The tool is based on the initial elections taken by the designer. Then and successively, it is shaped on the form of different operative steps, criteria, sub‐criteria and possibilities which respond to a different progress in the definition of the building construction system. In order to inform about the stage of the decision and the definition reached by the project in every particular case, a range of operative sheets are proposed. Additionally, the system allows the connection with other reviewing methods for building projects. The aim of this last possibility is to encourage the reflection on standardization of the associated risks to the building system itself and its future performance. The tool provides a helping system to be used during the decision making process for a multilayer façade design. It minimizes the arbitrariness and offers a qualification previous to the quantification that will be done with the development of the construction details and their bill of quantities, that in subsequent project stages will be executed. At the same time, the systematization of the mentioned decision making during the design phase, can be found as a checking system in the different stages of the decision making design process and in the different stages of the building envelope definition.

A Vision-based Quadrotor Multi-robot Solution for the Indoor Autonomy Challenge of the 2013 International Micro Air Vehicle Competition

This paper presents a completely autonomous solution to participate in the Indoor Challenge of the 2013 International Micro Air Vehicle Competition (IMAV 2013). Our proposal is a multi-robot system with no centralized coordination whose robotic agents share their position estimates. The capability of each agent to navigate avoiding collisions is a consequence of the resulting emergent behavior. Each agent consists of a ground station running an instance of the proposed architecture that communicates over WiFi with an AR Drone 2.0 quadrotor. Visual markers are employed to sense and map obstacles and to improve the pose estimation based on Inertial Measurement Unit (IMU) and ground optical flow data. Based on our architecture, each robotic agent can navigate avoiding obstacles and other members of the multi-robot system. The solution is demonstrated and the achieved navigation performance is evaluated by means of experimental flights. This work also analyzes the capabilities of the presented solution in simulated flights of the IMAV 2013 Indoor Challenge. The performance of the CVG UPM team was awarded with the First Prize in the Indoor Autonomy Challenge of the IMAV 2013 competition.

Structure of the Ets-1 pointed domain and mitogen-activated protein kinase phosphorylation site

The Pointed (PNT) domain and an adjacent mitogen-activated protein (MAP) kinase phosphorylation site are defined by sequence conservation among a subset of ets transcription factors and are implicated in two regulatory strategies, protein interactions and posttranslational modifications, respectively. By using NMR, we have determined the structure of a 110-residue fragment of murine Ets-1 that includes the PNT domain and MAP kinase site. The Ets-1 PNT domain forms a monomeric five-helix bundle. The architecture is distinct from that of any known DNA- or protein-binding module, including the helix-loop-helix fold proposed for the PNT domain of the ets protein TEL. The MAP kinase site is in a highly flexible region of both the unphosphorylated and phosphorylated forms of the Ets-1 fragment. Phosphorylation alters neither the structure nor monomeric state of the PNT domain. These results suggest that the Ets-1 PNT domain functions in heterotypic protein interactions and support the possibility that target recognition is coupled to structuring of the MAP kinase site.

Molecular characterization of the mycobacterial heparin-binding hemagglutinin, a mycobacterial adhesin

Although it generally is accepted that the interaction of Mycobacterium tuberculosis with alveolar macrophages is a key step in the pathogenesis of tuberculosis, interactions with other cell types, especially epithelial cells, also may be important. In this study we describe the molecular characterization of a mycobacterial heparin-binding hemagglutinin (HBHA), a protein that functions as an adhesin for epithelial cells. The structural gene was cloned from M. tuberculosis and bacillus Calmette–Guérin, and the sequence was found to be identical between the two species. The calculated Mr was smaller than the observed Mr when analyzed by SDS/PAGE. This difference can be attributed to the Lys/Pro-rich repeats that occur at the C-terminal end of the protein and to a putative carbohydrate moiety. Glycosylation of HBHA appears to protect the protein from proteolytic degradation, which results in the removal of the C-terminal Lys/Pro-rich region responsible for binding of HBHA to sulfated carbohydrates. Evidence suggests that glycosylation is also important for HBHA-mediated hemagglutination and for certain immunologic properties of the protein. Finally, the absence of a signal peptide in the coding region of HBHA raises the possibility that this protein is not secreted via the general secretion pathway.

NEAT - A New, Evolutive API and Transport-Layer Architecture for the Internet

ACKNOWLEDGMENTS This work has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 644334 (NEAT). The views expressed are solely those of the authors.

The protein product of the het-s heterokaryon incompatibility gene of the fungus Podospora anserina behaves as a prion analog

The het-s locus of Podospora anserina is a heterokaryon incompatibility locus. The coexpression of the antagonistic het-s and het-S alleles triggers a lethal reaction that prevents the formation of viable heterokaryons. Strains that contain the het-s allele can display two different phenotypes, [Het-s] or [Het-s*], according to their reactivity in incompatibility. The detection in these phenotypically distinct strains of a protein expressed from the het-s gene indicates that the difference in reactivity depends on a posttranslational difference between two forms of the polypeptide encoded by the het-s gene. This posttranslational modification does not affect the electrophoretic mobility of the protein in SDS/PAGE. Several results suggest a similarity of behavior between the protein encoded by the het-s gene and prions. The [Het-s] character can propagate in [Het-s*] strains as an infectious agent, producing a [Het-s*] → [Het-s] transition, independently of protein synthesis. Expression of the [Het-s] character requires a functional het-s gene. The protein present in [Het-s] strains is more resistant to proteinase K than that present in [Het-s*] mycelium. Furthermore, overexpression of the het-s gene increases the frequency of the transition from [Het-s*] to [Het-s]. We propose that this transition is the consequence of a self-propagating conformational modification of the protein mediated by the formation of complexes between the two different forms of the polypeptide.

Identification of a small, very acidic constitutive nucleolar protein (NO29) as a member of the nucleoplasmin family

We report the discovery and molecular characterization of a small and very acidic nucleolar protein of an SDS/PAGE mobility corresponding to Mr 29,000 (NO29). The cDNA-deduced sequence of the Xenopus laevis protein defines a polypeptide of a calculated molecular mass of 20,121 and a pI of 3.75, with an extended acidic region near its C terminus, and is related to the major nucleolar protein, NO38, and the histone-binding protein, nucleoplasmin. This member of the nucleoplasmin family of proteins was immunolocalized to nucleoli in Xenopus oocytes and diverse somatic cells. Protein NO29 is associated with nuclear particles from Xenopus oocytes, partly complexed with protein NO38, and occurs in preribosomes but not in mature ribosomes. The location and the enormously high content of negatively charged amino acids lead to the hypothesis that NO29 might be involved in the nuclear and nucleolar accumulation of ribosomal proteins and the coordinated assembly of pre-ribosomal particles.

cDNA cloning of Batis maritima methyl chloride transferase and purification of the enzyme

Methyl chloride transferase catalyzes the synthesis of methyl chloride from S-adenosine-l-methionine and chloride ion. This enzyme has been purified 2,700-fold to homogeneity from Batis maritima, a halophytic plant that grows abundantly in salt marshes. The purification of the enzyme was accomplished by a combination of ammonium sulfate fractionation, column chromatography on Sephadex G100 and adenosine-agarose, and TSK-250 size-exclusion HPLC. The purified enzyme exhibits a single band on SDS/PAGE with a molecular mass of approximately 22.5 kDa. The molecular mass of the purified enzyme was 22,474 Da as determined by matrix-associated laser desorption ionization mass spectrometry. The methylase can function in either a monomeric or oligomeric form. A 32-aa sequence of an internal fragment of the methylase was determined (GLVPGCGGGYDVVAMANPER FMVGLDIXENAL, where X represents unknown residue) by Edman degradation, and a full-length cDNA of the enzyme was obtained by rapid amplification of cDNA ends–PCR amplification of cDNA oligonucleotides. The cDNA gene contains an ORF of 690 bp encoding an enzyme of 230 aa residues having a predicted molecular mass of 25,761 Da. The disparity between the observed and calculated molecular mass suggests that the methylase undergoes posttranslational cleavage, possibly during purification. Sequence homologies suggest that the B. maritima methylase defines a new family of plant methyl transferases. A possible function for this novel methylase in halophytic plants is discussed.

Cloning of the cDNA encoding the large subunit of human RNase HI, a homologue of the prokaryotic RNase HII

Two RNases H of mammalian tissues have been described: RNase HI, the activity of which was found to rise during DNA replication, and RNase HII, which may be involved in transcription. RNase HI is the major mammalian enzyme representing around 85% of the total RNase H activity in the cell. By using highly purified calf thymus RNase HI we identified the sequences of several tryptic peptides. This information enabled us to determine the sequence of the cDNA coding for the large subunit of human RNase HI. The corresponding ORF of 897 nt defines a polypeptide of relative molecular mass of 33,367, which is in agreement with the molecular mass obtained earlier by SDS/PAGE. Expression of the cloned ORF in Escherichia coli leads to a polypeptide, which is specifically recognized by an antiserum raised against calf thymus RNase HI. Interestingly, the deduced amino acid sequence of this subunit of human RNase HI displays significant homology to RNase HII from E. coli, an enzyme of unknown function and previously judged as a minor activity. This finding suggests an evolutionary link between the mammalian RNases HI and the prokaryotic RNases HII. The idea of a mammalian RNase HI large subunit being a strongly conserved protein is substantiated by the existence of homologous ORFs in the genomes of other eukaryotes and of all eubacteria and archaebacteria that have been completely sequenced.

The orientation of the AP-1 heterodimer on DNA strongly affects transcriptional potency

Activation of gene transcription in eukaryotes requires the cooperative assembly of an initiation complex containing many protein subunits. The necessity that these components contact each other and the promoter/enhancer in defined ways suggests that their spatial arrangement might influence the activation response. Indeed, growing evidence indicates that DNA architecture can profoundly affect transcriptional potency. Much less is known about the influence of protein architecture on transcriptional activation. Here, we examine the architectural dependence of activator function through the analysis of matched pairs of AP-1•DNA complexes differing only in their orientation. Mutation of a critical Arg residue in the basic-leucine zipper domain of either Fos or Jun yielded single point-mutant heterodimers that bind DNA in a single defined orientation, as determined directly by native chemical ligation/affinity cleavage; by contrast, the corresponding wild-type protein binds DNA as a roughly equal mixture of two isomeric orientations, which are related by subunit interchange. The stereochemistry of the point-mutant heterodimers could be switched by inversion of a C•G base pair in the center of the AP-1 site, thus providing access to both fixed orientational isomers. Yeast reporter gene assays consistently revealed that one orientational isomer activates transcription at least 10-fold more strongly than the other. These results suggest that protein architecture, especially the spatial relationship of the activation domain to the promoter, can exert a powerful influence on activator potency.

Tissue phenotype depends on reciprocal interactions between the extracellular matrix and the structural organization of the nucleus

What determines the nuclear organization within a cell and whether this organization itself can impose cellular function within a tissue remains unknown. To explore the relationship between nuclear organization and tissue architecture and function, we used a model of human mammary epithelial cell acinar morphogenesis. When cultured within a reconstituted basement membrane (rBM), HMT-3522 cells form polarized and growth-arrested tissue-like acini with a central lumen and deposit an endogenous BM. We show that rBM-induced morphogenesis is accompanied by relocalization of the nuclear matrix proteins NuMA, splicing factor SRm160, and cell cycle regulator Rb. These proteins had distinct distribution patterns specific for proliferation, growth arrest, and acini formation, whereas the distribution of the nuclear lamina protein, lamin B, remained unchanged. NuMA relocalized to foci, which coalesced into larger assemblies as morphogenesis progressed. Perturbation of histone acetylation in the acini by trichostatin A treatment altered chromatin structure, disrupted NuMA foci, and induced cell proliferation. Moreover, treatment of transiently permeabilized acini with a NuMA antibody led to the disruption of NuMA foci, alteration of histone acetylation, activation of metalloproteases, and breakdown of the endogenous BM. These results experimentally demonstrate a dynamic interaction between the extracellular matrix, nuclear organization, and tissue phenotype. They further show that rather than passively reflecting changes in gene expression, nuclear organization itself can modulate the cellular and tissue phenotype.

The active oligomeric state of the minimalistic influenza virus M2 ion channel is a tetramer

The influenza A virus M2 integral membrane protein is an ion channel that permits protons to enter virus particles during uncoating of virions in endosomes and also modulates the pH of the trans-Golgi network in virus-infected cells. The M2 protein is a homo-oligomer of 97 residues, and analysis by chemical cross-linking and SDS/PAGE indicates M2 forms a tetramer. However, a higher order molecular form is sometimes observed and, thus, it is necessary to determine the active form of the molecule. This was done by studying the currents of oocytes that expressed mixtures of the wild-type M2 protein (epitope tagged) and the mutant protein M2-V27S, which is resistant to the inhibitor amantadine. The composition of mixed oligomers of the two proteins expressed at the plasma membrane of individual oocytes was quantified after antibody capture of the cell surface expressed molecules and it was found that the subunits mixed freely. When the ratio of wild-type to mutant protein subunits was 0.85:0.15, the amantadine sensitivity was reduced to 50% and for a ratio of 0.71:0.29 to 20%. These results are consistent with the amantadine-resistant mutant being dominant and the oligomeric state being a tetramer.

Interaction of the Doa4 Deubiquitinating Enzyme with the Yeast 26S Proteasome

The Saccharomyces cerevisiae Doa4 deubiquitinating enzyme is required for the rapid degradation of protein substrates of the ubiquitin–proteasome pathway. Previous work suggested that Doa4 functions late in the pathway, possibly by deubiquitinating (poly)-ubiquitin-substrate intermediates associated with the 26S proteasome. We now provide evidence for physical and functional interaction between Doa4 and the proteasome. Genetic interaction is indicated by the mutual enhancement of defects associated with a deletion of DOA4 or a proteasome mutation when the two mutations are combined. Physical association of Doa4 and the proteasome was investigated with a new yeast 26S proteasome purification procedure, by which we find that a sizeable fraction of Doa4 copurifies with the protease. Another yeast deubiquitinating enzyme, Ubp5, which is related in sequence to Doa4 but cannot substitute for it even when overproduced, does not associate with the proteasome. DOA4-UBP5 chimeras were made by a novel PCR/yeast recombination method and used to identify an N-terminal 310-residue domain of Doa4 that, when appended to the catalytic domain of Ubp5, conferred Doa4 function, consistent with Ubp enzymes having a modular architecture. Unlike Ubp5, a functional Doa4-Ubp5 chimera associates with the proteasome, suggesting that proteasome binding is important for Doa4 function. Together, these data support a model in which Doa4 promotes proteolysis through removal of ubiquitin from proteolytic intermediates on the proteasome before or after initiation of substrate breakdown.