El centro comercial : una teoría tipológica : claves para el reconocimiento de una arquitectura ignorada

Existe en el panorama edificado un patrimonio construido que se reconoce como Centro Comercial. Un conjunto entendido, en sentido genérico, como familia arquitectónica que tiene características propias y específicas que la identifican. El objeto de la presente tesis doctoral consiste en argumentar que este conjunto constituye un nuevo tipo en el panorama de las tipologías arquitectónicas. Un tipo con entidad propia, que se conecta a una forma diferente de entender la idea de modelo. Un concepto que va más allá de la consideración tradicional del término. Modelo virtual. Este tipo, que surge de una estructura teórica que hemos denominado teoría tipológica, se constituye en una herramienta más para el estudio y el desarrollo proyectual de los espacios arquitectónicos, tanto del propio Centro Comercial como del conjunto de la disciplina arquitectónica, como referencia legítima. El presente trabajo de tesis se inicia con un bloque introductorio denominado Método. Definimos en él una metodología que hemos llamado emocional. Trata de la oportunidad de la tesis. Del porqué de un título que recoge la palabra ignorada. Del interés que suscita el asunto en el contexto del momento presente. Oportunidad e interés en base a una vida profesional dedicada al mundo del Centro Comercial y a la importancia del patrimonio elegido como objeto de estudio. También ha sido un aliciente detectar como las planificaciones del territorio y de los ámbitos de las relaciones colectivas no han sido capaces de integrar un resultado satisfactorio. Quizás por no considerar la complejidad de sus muchas facetas. En consecuencia, el texto busca la esencia del Centro Comercial como soporte para la crítica de su impacto en los nuevos escenarios de relación que la sociedad y el entorno físico imponen. Ámbitos donde los mecanismos históricos del asociacionismo tradicional han dado paso a otros, como el Centro Comercial, de exaltación del individualismo, pero demandados por una sociedad que se identifica con ellos. Espacios que, con Galbraith, existen por de la perversidad intrínseca del binomio consumo-producción que inducen la perplejidad. Ésta pasa a formar parte de la esencia del nuevo espacio comercial, como quedó de manifiesto en el Congreso de Minnesota de 1997, sobre el Centro Comercial. Una sociedad que ha girado hacia el logro material en términos de culto, ocasionando creciente valor significante del consumo. Razón última de la humanidad al decir de Rem Koolhaas. Culto que desemboca en la urgencia de alcanzar niveles de estatus y de identificación con el grupo. Dos parámetros que marcan la relación con el otro. Relación de comparación que excita la necesidad de posesión de objetos que llevaban a recrear en el consumidor la ilusión de ser especiales, de no pasar desapercibidos. El producto de consumo, el objeto, se eleva a la consideración de valor social. En el Centro Comercial se venden valores. Marketing de valores. El deseo del individuo, no la necesidad, queda involucrado en el proceso. La oportunidad y el interés de este estudio surgen además para aclarar el significado de un espacio que sirve al consumidor y a su entorno. Un significado que alcanza sentido, entre otros, por la aparición de un nuevo contexto tras la Segunda Guerra Mundial. Suburbanización, automóvil, nivel de vida, un nuevo papel de la mujer. La Era del Consumo. Una era que genera la paradoja de un individuo con autodeterminación y autosuficiencia crecientes, en un mundo cada vez más condicionado y controlado por dinámicas de ofertas mediatizadas e ilimitadas. La arquitectura en este contexto ha de juzgarse por su relación con un hombre contemporáneo que camina hacia una progresiva excepcionalidad. Cerramos la presentación justificando el término ignorada que aparece en el Título de la Tesis. Subrayamos la escasa existencia de estudios estructurados que relacionen Centro Comercial y arquitectura. Realidad que se constata partiendo del libro de Pevsner, Historia de las Tipologías Arquitectónicas, de 1976. Analizamos también la aceptación que el Centro Comercial, valorado en algunos ámbitos como arquitectura populista. Primer Capítulo, los antecedentes. Se propone un recorrido histórico por la arquitectura comercial de todos los tiempos. El Documento busca concretar las características de la arquitectura comercial a lo largo de la historia para determinar relaciones entre ella y el Centro Comercial. Estas correspondencias van a permitir contestar la pregunta retórica que nos hacemos al inicio del Capítulo: ¿Es la arquitectura del Centro Comercial una arquitectura subsidiaria, o tiene personalidad propia independiente de la del mundo comercial? Con Zevi40, queremos dejar constancia que la relación arquitectura-historia es imprescindible en la tarea proyectual. Un análisis novedoso solo es posible desde la búsqueda de unas raíces auténticas. Así mismo, con Guy de Maupassant, estamos convencidos que “La arquitectura, a través de los siglos, ha tenido el privilegio de dar un símbolo a cada una de las épocas, de resumir con un pequeñísimo número de monumentos típicos, el modo de pensar, de soñar de una raza y de una civilización”. Este recorrido se inicia interpretando la arquitectura comercial minoica. Llegaremos, paso a paso, hasta el siglo XX, los grandes itinerarios comerciales y el Centro Comercial. Se descubren una serie de invariantes que permiten comparar y extraer conclusiones. Resulta novedoso constatar que el Centro Comercial nace para dar respuesta al hombre contemporáneo en su afán consumista. También lo es la rápida implantación y evolución del Centro Comercial en un corto periodo de sesenta años frente al ritmo sosegado de otras soluciones comerciales. Novedad es ver como el comercio y sus arquitecturas nacen abiertos, bajo tenderetes y el Centro Comercial se presenta cerrado. Así mismo, las referencias sacras constituyen un elemento de novedad para la reflexión, en un contexto materialista. Y tantas otras. En Minos, la óptica comercial ofrece otra visión de su legendaria cultura. ¿Palacio o plataforma logística?, gestión centralizada del intercambio, ¿vida pública y vida privada? Así, hasta llegar al siglo XIX. Sus las galerías y sus pasajes acristalados concebidos en primera instancia como medida de recomposición urbana. Espacio entendido desde lo público-privado, desde lo interior-exterior, desde el dentro-afuera, desde lo cerrado-abierto. Con los nuevos mercados de abastos, representan una revolución en el ámbito de la funcionalidad, de la máquina, de la gestión moderna y de las relaciones del ciudadano con la ciudad apoyado en base al intercambio de bienes de consumo. El Centro Comercial es heredero de esta transformación. El Gran Almacén, por otra parte, es el reflejo de otra gran revolución. La que va ligada a la producción y comercialización en masa, las comunicaciones, el precio fijo y el aumento del nivel de vida. El Centro Comercial reinterpreta estas situaciones insertándolas en un nuevo modelo de gestión. En el siglo XX, maestro en técnicas de venta, aparece el hipermercado. El Híper, con su carga de pedagogía, se incorpora al esquema orgánico del Centro Comercial. La tienda en si misma constituye la pieza base de dicho puzle orgánico. Es en esta época cuando la tienda empieza su despegue autónomo como arquitectura, aportando su experiencia. Tras ello, llegamos a las grandes rutas comerciales, que proponemos como metáfora del shopping. Cerramos el capítulo concluyendo con Eugenio Ferrer que “si establecemos una relación entre el espacio y el capitalismo, entonces podemos inferir que los espacios del consumo de masas (ECM) son configuraciones nuevas respecto al pasado (...), pero el sistema que lo introduce no lo es del todo”. Segundo Capítulo. Búsqueda de claves que permitan el reconocimiento del Centro Comercial. Llegados a este punto y con la perspectiva del tiempo cabe preguntarnos, ¿qué entendemos pues por Centro Comercial? ¿Cómo lo percibimos? Abordamos ahora el problema de la percepción del Centro Comercial y su significado. Además de constituirse en símbolo, referencia siempre presente, la eficacia del Shopping es una de las principales causas de su poder de atracción. El Centro Comercial resulta de la síntesis de la revolución del consumo y de la revolución de la gestión. Espacio eficaz del entretenimiento como destino. El usuario resuelve su vida cotidiana dentro de un hábitat que considera propio y que se entiende como el lugar hiperbólico de la transacción comercial. Espacio de la abundancia para el disfrute. Una arquitectura involucrada en esta eficacia. Su sentido del lugar no es otro que el Shopping, que se desenvuelve de forma análoga en todas partes. El hogar del consumidor. Las nuevas catedrales. Las catedrales del consumo. Destino místico. Lugar de peregrinación para el consumidor fervoroso. Espacio sagrado que integra al usuario en la cultura dominante. Cultura del consumo. Templos, donde el tiempo ha perdido su sentido. Paraíso. Un espacio donde la altura, la luz natural, la dimensión general refuerzan el carácter sacro de un espacio para una nueva religión laica. Un espacio seguro, protegido que nos acerca a ideas como la de útero materno, con su carga de calidez y de significación erótica aplicado al encuentro compra-comprador y, ligado a ello, la literatura especializada habla del Centro Comercial como nave espacial hiperesterilizada o de agujero negro que absorbe la energía cultural. Más allá, la simulación, donde se percibe un simulacro de ciudad. Simulacro coherente con todo lo que el Shopping desencadena a su alrededor. El lugar de los sueños, de la fantasía. Aquí los productos se han metamorfoseado en fetiches, en significados. Televisión tridimensional, donde el usuario actúa guiado por una pulsión similar al zapping. Espacio lúdico de la fascinación por comprar o de imaginar que se compra. Espacio de simulaciones que llevan a la ensoñación. Un nuevo lugar que sustituye al espacio cotidiano, con el señuelo de la protección, en un contexto imaginariamente público. Espacio de la hiperrealidad donde no se distingue la realidad de la fantasía, donde tras episodios de confusión y desconcierto, se borran las fronteras con lo imaginario. Espacio mágico, escenario del gran espectáculo del consumo, controlado milimétricamente, al modo de la visita a un gran monumento, que ha de sobrevivir a los tiempos para dar testimonio de nuestro momento. Un icono, un símbolo que transmite un mensaje, que solo el consumidor es capaz de interpretar. Una agitada mezcla, sin precedentes, de percepciones que hablan de perplejidad y asombro ante el fenómeno del Centro Comercial, su espacio y las reacciones del hombre contemporáneo. ¿Cuáles serían las claves que nos permitirían reconocer la calidad esencial de un Centro Comercial, en esta concurrencia de perplejidades? Primero, la función de servicio. Un espacio donde ocurren muchas más cosas que lo obvio, que la compra. Un edificio que se involucra con el entorno de la mano de lo inesperado, la sorpresa y las expectativas. Esta vocación de servicio conecta Centro Comercial y naturaleza arquitectónica. Función que sugiere percepciones ligadas a la experiencia de compra. Organismo que vive y late al unísono con su visitante, colocándose al servicio de sus necesidades, de su afán de consumo, del que vive. Segundo, la comunidad servida. El Centro comercial sirve a una sociedad concreta. La sociedad consumista. Una nueva sociedad que se identifica con el edificio desarrollando un sentido de comunidad al nivel de sus deseos. Esta comunidad que se configura a su alrededor, constituye el activo más poderoso para el éxito de su realidad cotidiana y de su futuro. Tercero, un compromiso de carácter holístico. La economía de la experiencia aplicada al afán consumista de una sociedad identificada con su Centro, da lugar a una experiencia holística planificada. Diseño emocional. Colaboración para el éxito de un conjunto de establecimientos comerciales que participan en la aventura espacio-comercial del Centro Comercial. Ellos son los inductores primarios del consumo. Pero esta colaboración tiene su culminación en la amplificación del mensaje, como un inmenso altavoz, que proviene de la unidad configurada por todos ellos. El reflejo de esta amplificación de mensajes, desde la poderosa unidad constituida, es el aumento de la rentabilidad, fin último de la operación. Cuarto, la forma a través de una identificación de carácter gestáltico. Desde la lectura gestáltica que hacemos de la unidad holística, se advierte una poderosa capacidad de comunicación del sistema con su contexto. Centro y entorno se tornan entonces cómplices que complementan sus realidades. El Centro Comercial, arquitectura estructurada como sistema se percibe –ha de percibirse- como forma unitaria que procede de una mirada de raíz gestáltica que continuamente la recompone desde una óptica espacial y física, ligada a la experiencia individual. Esta unidad formal, más allá de la forma real, se constituye en esencia de de su arquitectura. Quinto, el Centro Comercial como sistema. Un sistema soportado por la Teoría General de Sistemas. La consideración del Centro Comercial como sistema es consecuencia de su estructura holística. El todo no se comporta como la suma de las partes y estas no lo hacen como lo harían en solitario. De aquí surge la necesidad de diálogo permanente entre la comercialización –proceso de incorporación de partes- y su traducción al mundo del diseño –proceso de articular arquitectónicamente las partes. Como sistema así configurado, el Centro Comercial se inserta en el paradigma contemporáneo, lo que genera realidades duales que no son excluyentes y reacciones de perplejidad e incertidumbre que el sistema corrige con su capacidad de autorregulación. Aparece también el espectador cuántico, el visitante, el consumidor, que interactúa con el sistema. Desde las herramientas que nos aporta la idea de sistema complejo, afrontamos el Mix Comercial -en definitiva la eficaz localización de las piezas en orden a sus relaciones y al organismo resultante- y su incidencia en la arquitectura que estamos concibiendo. Una arquitectura impredecible por lo mutable, que se erige en reto de la operación y del diseñador. Diseño que, de la mano del concepto de sistema se convierte en herramienta a mayor gloria de la operación global. El debate del estilismo no será más que el resultado del análisis en busca del éxito de esta operación. Sexto, una arquitectura de la negociación. Negociación como mecanismo proyectual y como resultado. La solución de proyecto nunca resulta evidente en el Centro Comercial como consecuencia de lo imprevisible del propio proceso de configuración. Su concreción solo puede ser fruto del compromiso de todos los agentes por conseguir el objetivo de la operación. Esto se consigue desde el equilibrio de intereses. Comerciales y de diseño. Un compromiso con la negociación y una negociación íntimamente ligada a la coordinación. Séptimo, el espacio y el tiempo. El debate espacio-tiempo condiciona y estructura la percepción del Centro Comercial. Introducimos conceptos como cronotopo –el instante y el lugar donde ocurre algo- y paradoja –incoherencia de la relación causa efecto- que sitúan el vínculo entre el tiempo y el espacio del Centro Comercial en un contexto de Shopping. En el discurrir paralelo del tiempo histórico –el tiempo de fuera- y del tiempo interior, el de dentro del Centro Comercial –tiempo presente o intemporalidad-, se produce el triunfo social del Centro Comercial que se traduce en haber sabido resolver en el espacio y en el tiempo las paradojas postmodernas del hombre contemporáneo. Octavo, de lo global. Globalidad que no es ajena a lo local. Una arquitectura que insertada en la dinámica de una economía de ámbito mundial, refleja las contradicciones que ella impone, fundamentalmente en los procesos de inclusión y exclusión, afectando de manera decisiva al debate de lo local, que el Centro Comercial debe incorporar como herramienta ineludible de reconocimiento. Terminamos el capítulo segundo manifestando como estas ocho claves, asumidas en su conjunto, confirman que el Centro Comercial puede aparecer como un todo conceptual cohesionado, pasando a formar parte de una familia arquitectónica coherente, cuya estructura funcional somos capaces de establecer. El Capítulo Tercero presenta con detalle la figura del arquitecto Víctor Gruen, creador reconocido del moderno Centro Comercial. Presentamos su trayectoria profesional observando como las diferentes claves analizadas en capítulos anteriores van apareciendo de manera natural a lo largo de ella "Victor Gruen may well have been the most influential architect of the twentieth century." Malcolm Gladwell. The Terrazo Jungle. Fifty Years Ago, The Mall Was Born. America Never Would Be The Same. 2004. In The New Yorker. Hombre complejo, conflictivo. Las paradojas a lo largo de su carrera fueron notables. Sin embargo siempre apareció como un hombre recto. Garret Eckbo, reconocido paisajista y colaborador de Gruen en el Centro Comercial peatonal del Centro urbano de Fresno, California, se asombraba de que alguien como Gruen hubiese sido capaz de combinar cortesía y humanidad en la carrera de ratas (sic) en la que se convirtió el universo de la comercialización americana y mantener la integridad542. Philip Johnson, en 1962, ponderando las muchas habilidades de Gruen manifestó que no estaba seguro si alguno de ellos, arquitectos artistas, hubiera sido capaz de hacer lo que Gruen hizo. A más, sobre Gruen, manifestó lo siguiente: "... El va más allá de la creación de un bello edificio. En jugar con la gente y sugerir lo que tienen que hacer, es un maestro. Y obtiene buenos resultados como hace la escultura. El suyo es un arte cívico, un sentido cívico.... Él es capaz de sentarse y poner cosas juntas. No es pomposo ni vano. Yo no me reuniría con él para hablar del diseño (de edificios). Víctor siente que cuando se habla de diseño se está ignorando todo el contexto... Su arquitectura es poderosamente limpia, no vuela la fantasía. Pero cuando te haces con su complejidad, ves que has descubierto algo más allá del diseño.... No puedes decir que haya alguien como él. La arquitectura tiene la suerte de tenerle como arquitecto..." Philip Johnson. Article in Fortune Review. 1962. El Documento de tesis cierra la visión de Gruen realizando un recorrido por los diferentes capítulos de su libro esencial, Shopping Towns Usa: The Planning of Shopping Centers. Solo su índice resulta un monumento al proceso de gestación del Centro Comercial. El libro, aquí simplemente mostrado en su estructura básica como un silencioso testigo, es la referencia canónica los Centros Comerciales contemporáneos, desde su aparición. . El Cuarto Capítulo del Documento de tesis es una recapitulación del trabajo anterior, en el que se sintetizan los conceptos de función y estilo relacionados con el Centro Comercial, se define en qué consiste cada uno de ellos y como, a partir de ahí, podemos afirmar que nos encontramos ante un tipo arquitectónico nuevo en el panorama de la disciplina arquitectónica. Terminamos el Capítulo integrando el Centro Comercial con un cuerpo teórico de referencias que se remiten a un tipo arquitectónico concreto y particular, acogiendo su singularidad como fenómeno arquitectónico autónomo. Como Conclusión de la tesis, resultado de todo lo anterior es decir, como consecuencia de la integración de un torrente de percepciones e intuiciones en un cuerpo teórico de referencias, deducidas de la existencia de unas claves que estructuran y penetran la esencia del singular modelo estudiado, haciéndolo detectable y seductor, resultan las características de un tipo arquitectónico con entidad propia que ordena, orienta y supedita la realidad y la existencia de esta nueva arquitectura. Una arquitectura nunca antes definida como tal, en el panorama tipológico de la disciplina. Teoría tipológica para una nueva arquitectura, que hemos ido proponiendo a lo largo del trabajo y que es coherente con los diferentes parámetros que se han analizado. Un conjunto edificado que, desde el estudio de sus claves esenciales y de sus invariantes perceptibles, aparece ahora más cercano, más familiar. Tanto que es posible destilar desde este conocimiento cercano e íntimo, una síntesis útil como referencia proyectual y como referencia para las grandes cuestiones que preocupan al discurrir del debate arquitectónico y sus ideas. El debate de la disciplina. El objeto de esta tesis, que consistía en establecer que el conjunto edificado que conocemos como Centro Comercial se constituye en un nuevo tipo en el panorama de las tipologías arquitectónicas, entendido el Centro Comercial en sentido genérico como familia arquitectónica con características propias y específicas que la hacen autónoma y reconocible, queda a nuestro juicio argumentado y justificado. ABSTRACT Within the frame of the built heritage there is a construction that is recognized as Shopping Center. An ensemble understood as an architectural family with its own specific recognizable characteristics. This thesis aims to explain that this building complex constitutes a new type in the panorama of architectural typology. This typology, with its singular identity, is connected to a way of understanding the idea of the model beyond an orthodox conception of the term understood as virtual model. This typology comes from a theoretical structure that we called typological theory, and it serves as yet another tool to reference the study and development design of the architectural spaces. In this first section, the Method, we emotionally explore the opportunities of this thesis. Why this typology has been ignored and the interest this work has in the present moment. An opportunity and an interest explained from an experience of a life dedicated to the world of Shopping Centers. The text then introduces the need for a rigorous knowledge of the Shopping Center’s essence in order to understand its impact in the frame of a new society and a new physical environment. A frame time where the historical mechanisms of association of civic community have given way to other gathering spaces like the Shopping Center, which encourages individualism, but is demanded by a society that relates to them. Spaces that, according to Galbraith, are a result of the intrinsic perversity of the unstoppable movement of the wheel of consumption-production. A society that has turned to worship of material achievement. Worship that provokes the appearance of an increasing value of consumption, according to Koolhaas, the only goal of humanity. Worship that ends in the need to reach certain status levels in the plane of a permanent comparison where the need of possession excites the consumer and gives them the illusion of being special. The product of consumption rises up to consideration of social values, entering a dynamic of marketing values, not only objects, but the desire of the individual remains. The study appears also to clarify the meaning of a space that serves the consumer and its context. A meaning that makes sense with the appearance of the suburbanization, the massive utilization of the car, the increase of living standards and the new role of women in the society after the Second World War, giving rise to the Age of Consumption. A world now determined and controlled by media and unlimited offers, where it’s necessary to place them in the context of the ordinary. An architecture that has to be judged precisely for its relation with this specific contemporary man. This section ends justifying the term ignored that appears in the Title of the Thesis, considering it in relation with the lack of studies structured about the Shopping Center and its architecture, drawing from Pevsner's work, A history of building types, 1976. Finally, the Shopping Center is analyzed with the most critical of thoughts, which considers it as populist architecture. The First Chapter, Precedents, proposes an historical tour of the commercial architecture throughout history. The Document looks to place on record the characteristics of the commercial architecture to set the connection between them and the Mall itself. These correspondences are going to allow us to answer the rhetorical question: is it the architecture of the Shopping Center a subsidiary architecture, or does it have its own personality independent from that of the commercial world? The reason of this historical search, citing Bruno Zevi, is that it is indispensable to establish the relationship between architecture and history, understating that an analysis is only possible when researching for their roots. Moreover, according to Guy de Maupassant, we are sure that architecture has had the privilege, across the centuries, of symbolizing as it were each age(…), through the harmony of lines and the charm of ornamentation all the grace and grandeur of an epoch. This historical reading, inseparable from a consistent design action, begins interpreting the commercial architecture of the Minoan to the 20th Century. Though this analysis of the big commercial itineraries and the study of the Shopping Center itself. A reading where we have found a series of constants that make it possible to draw conclusions from this comparison. The Mall appears to give response to the needs of a consumerist society. Comparing to the calm pace of the evolution of other commercial solutions, it is relevant its fast implantation and evolution in a short period of sixty years. Though via different solutions, the commercial spaces are considered taking into account the public-private relation, the interior-exterior, the inside-out, the closed-opened. Through that, the 19th century galleries and the food markets represent a revolution in functionality, in the machine, the modern management and the relations of the citizen within the city. All of this, the Mall inheritor feels. Likewise, the Department Store is the reflection of another great revolution. Production and commercialization en-mass, communications, the fixed price and increase of the living standard. The Mall reinterprets these situations inserting them in a new model of management. Already in the 20th century Mall and mass technologies of sale, the hypermarket is enthusiastically incorporated into the configuration of this organic scheme, constituting the base of one of the models, the French, that will be highly developed in the European continent. The shop itself constitutes, on the other hand, the key piece that completes the puzzle of the Mall and is in this epoch when it starts taking off as architecture, has an autonomous character. After all this, finally, we come to the big commercial routes, which we propose as metaphor of the shopping. Citing Eugenio Ferrer we can conclude that “If we establish a relation between the space and the capitalism, then we can infer that the spaces of the consumption of masses (ECM) are new configurations with regard to the past (...), but the system that introduces it it is not completely” Now we arrive at this point and with the perspective of time it is necessary to ask us, what do we understand about the Shopping Center? How do we perceive it? The second Chapter approaches the problem of the perception of the Mall and from this it is possible to detect and to identify key drivers that orientate the architectural comprehension of the space. The efficiency of the Shopping Center is its main power of attraction. A world that has ensued from the synthesis of the revolution of consumption and management. An effective cavern-like place of entertainment where the user, the consumer, the postmodern man solves his daily life inside a considered habitat. The hyperbolic place of commercial transaction. An abundance of space, that makes us perceive it as destination of entertainment. An architecture has evolved this efficiency, where the sense of place is at one with the sense of the Shopping. The home of the consumer. The new cathedrals. The cathedral of consumption. The place of peregrination for the fervent consumer. A sacred space that integrates the user in the dominant culture. A temple, where time itself has stopped existing. In this paroxysm, an expression of the Garden of Eden or Paradise itself. A space where the height, the natural daylight and the spatial dimension reinforce the sacred character of a new lay religion. Another common perception is that of a protected area, which leads to metaphors and considerations that suggest the idea of maternal womb, with its weight of erotic meaning, referring to the encounter of the shopper making a purchase. The literature also tells us about its perception as a sterile space capsule, a black hole that absorbs all cultural energy. Likewise, a world simulation where a mock city is perceived at first instance. Consistent with all that shopping triggers inside. A city, a space conceived as a place of dreams, fantasy, where the products have been metamorphosed into fetishes. Entertaining a television, three-dimensional television, where the user acts guided by a drive similar to zapping. A play area where the latest fascination is in the act of buying. Space simulations that unite and transcend creating atmospheres that lead to reverie. A new space replacing the daily space with the lure of safe space in a public context. A hyper-reality space with reality and fantasy, where borders are erased with imaginary episodes of confusion and bewilderment, without distinction. The charm and fascination of a space that reads like magic. The magic of a space which is defined as stage extravaganza, the large theatre, the consumer surveys in the fine control mode in which you visit a national monument. The shopping center has to survive the times to be a testimony of our time. An icon, a symbol that conveys a message, the message reads ‘consumer’. In short, a Shopping Center is a mix of unprecedented insights that speak of a widespread phenomenon of bewilderment. Its space and the reactions of contemporary man unfold in it like a fish in water. What are the key aspects which allow us to recognize the essential quality of a shopping center in this concurrence of perplexities? First, we want to record a service function of space much deeper than the immediately obvious, i.e. a purchase occurs. A building that appears to be involved with the environment and its people from the hand of the unexpected circumstances; surprise and attention. And that, in turn, also involves the visitor beyond the purchase. This dedication to service closely links the mall with its architectural nature. It is not the function of a lifeless machine. It is a feature that suggests unsuspected perceptions linked to the purchase, which speaks of an organism that lives and breathes in unison with the visitor. Second, in addition to the vocation of service-oriented desire for consumption, the Mall environment serves a particular society - The consumer society. A new society which relates to building a sense of community developed to the level of their desires. This community also constitutes the most powerful asset to the success of the daily life of the Shopping Center. Third, we emphasize that the so called economy of the experience is combined with the consumer zeal of a company that is identified by the Shopping Center. It connects to form a holistic and planned experience. This experience takes shape in the entity that ensues from the premeditated association and synergy, in the sense of a collaboration for success. A set of concrete commercial and independent establishments, take part in the spatial and commercial adventure that is the Mall and they are the instigators of the consumption. This holistic behavior finds culmination in the amplification of a claim that becomes unitary, like an immense force that leads to an increased profitability to all the levels. Consummation is a reference of one human being overturned in an architecture assimilated into a legitimate, emotional design with stability. A holistic quality is born of the essence of the building - and by virtue of the Conditions of Alexander, Christopher Alexander, determines the system condition of the Shopping Center. Fourth, we propose to establish what character the Mall will form when joined with the concept of its typology. This is going to allow the architectural work to be formed. As a result of the holistic structure that we see, the Mall is perceived as a system whose parts have their own function, justifying their existence in the ecosystem. Across a gestalt there is a powerful capacity of communication between the system and its context. We visualize on the one that stands out our building, turning both, Center and environment, in accomplices of a few special relations who complement each other in his realities. This relationship within a complex and diverse environment gives the Mall a range of unique spatial perceptions, the result of disparate experiences, which because of its root origin of gestalt, are integrated into a unified and coordinated manner fully intelligible and organized. This is the final formal essence of the Shopping Center. We can conclude here that the Mall as architecture is a structured system and should be perceived as a unit both from a physical and spatial perspective as this is the essence of its architecture. Fifth, the Mall as a system. A system which is being supported by a broad theoretical corpus, the General Systems Theory, which offers sufficient methodology to descend into consideration and give an enlightened conclusion on the overall understanding of the Mall. Consideration of the Mall as a system is a result of its holistic structure. The whole does not behave like any of the parties and they do not behave the way they used to before belonging to the whole, because they inhibit many of their qualities to their advantage. It arises the need for an ongoing dialogue between marketing processes and its translation into the physical world, the design. The system generates multiple perceptions to be integrated into a body which is to be understood as unitary. As a system, the Mall is inserted into the contemporary paradigm, creating dual realities that are not exclusive and are reactions of uncertainty that the system be properly designed at all levels, faced with their ability to self-regulate. Likewise, considering the visitor, the customer, like the quantum spectator who interacts with the system permanently. Moreover, a complex system confronts us with the Commercial mix, the effective location of parts in order to relate to the body and its importance in the architecture we are conceiving. Unpredictable architecture, which stands as the challenge of the operation and the designer. Design that becomes the tool of the system to create success for the overall operation. The discussion of the styling is merely the result of analysis that also seeks the success of the system, i.e. the styling should send the right message for the environment to ensure its survival. Sixth, the idea of negotiation as an architecture project, a mechanism inherent to the status of the proposed system. The project solution is never evident at the Shopping Center because of the unpredictability of the process itself. It can only be the fruit of the commitment of all stakeholders to achieve the objective of the operation. This is achieved from the balance of interests, of commercial and design. A commitment to negotiation and a negotiation linked to coordination. The pursuit of stability is key, as instability is always present and constantly requires strategies to build the object you are configuring. Seventh, proposes space-time itself as a circumstance that determines and structures the perception of the Mall in a singular way. We introduce concepts as chronotope and paradox to help us place the relationship between time and space within the Mall in the context of shopping. A consequence of the parallel flow of historical time - the time outside - and the time inside the Mall, the big shopping center formula is precisely that of having the feeling of timelessness in the space. The social triumph of the mall is the ability to resolve in space and in time all postmodern paradoxes and, beyond that, of contemporary man, condensing into a small space and time an enormous amount of cultural symbols, often contradictory, but they attract the practice of consumerism. Eighth, global level. Globalization which doesn’t ignore the local level. Architecture that is inserted into the dynamics of a global economy, reflects the contradictions that it imposes, mainly in the processes of inclusion and exclusion. Inclusion and exclusion affect the debate of the local level, which the Mall must incorporate as an unavoidable tool of recognition. The eight fundamental principles, when applied as a whole, confirm that the built heritage, which corresponds to the general Mall idea, can be presented as a cohesive conceptual whole. This becomes part of a coherent architectural family, whose functional structures are able to be established. The Third chapter presents in a detailed way the figure of the architect Victor Gruen, recognized as the creator of the modern Mall. Studying his professional experience, it is shown how the different keys analyzed in previous chapters are appearing in a natural way. "Victor Gruen may well have been the most influential architect of the twentieth century." Malcolm Gladwell. The Terrazo Jungle. Fifty Years Ago, The Mall Was Born. America Never Would Be The Same. 2004. In The New Yorker. He was a complex, troubled man and the paradoxes along his career were notable. Nevertheless, always he appeared as a straight man. Garret Eckbo, the recognized landscape painter and collaborator of Gruen was astonished how Gruen had been capable of combining comity and humanity in this career of rates, into that the American commercialization turned, Johnson, in 1962, weighting many Gruen's skills demonstrated that he was not sure if anyone of them, architects artists, had been capable of doing what Gruen did. He goes beyond just the creation of a beautiful building. In playing on people and suggesting what they ought to do, he is a master. (…) his architecture is clean - hardly architecture, no flights of fancy. But when you get hold of its complexity, you've got something beyond the design... You can't say there's someone like him. Architecture is lucky to have him as an architect." Philip Johnson. Article in Fortune Review. 1962. The Document of the thesis closes with Gruen's vision of realizing a tour through the different chapters of his essential book, Shopping towns USA: The Planning of Shopping Centers. It’s mere index turns out to be a monument to the process of the gestation of the Mall. The book, simply acted in its basic structure as a silent witness, as the canonical reference for the contemporary Malls. The Fourth Chapter of the Document of the thesis is a recapitulation of the previous work, which synthesizes the concepts of function and style related to the Shopping Center, and clearly defines how they are defined so we can conclude that we have found an architectural new type in the panorama of the architectural discipline. Therefore, the Conclusion of the thesis integrates this development in a theoretical body of references that relate to an architectural specific and particular type, which receives the singularity of the Shopping Center as an architectural independent phenomenon as it has tried to demonstrate from the beginning of the work. To conclude, as a result of the integrative process and the development of the theoretical body of references, the essential characteristics of the order and concept of the architectural typology form the existence of a new architecture; architecture never before defined as such, in the theoretical typology of the discipline. A theoretical typology for a new architecture is proposed throughout the discussed research and forms a conclusion of the different parameters that have been analysed. As a building complex, from the study of the essential characteristics and of the perceptible constants, the typology is more clearly defined and thus, becomes a useful tool and precedent for the consideration of the discipline. The thesis then justifies how the building complex known as Shopping Center constitutes a new type of architectural typology.

Fluid-solid interaction in arteries incorporating the autoregulation concept in boundary conditions

In pre-surgery decisions in hospital emergency cases, fast and reliable results of the solid and fluid mechanics problems are of great interest to clinicians. In the current investigation, an iterative process based on a pressure-type boundary condition is proposed in order to reduce the computational costs of blood flow simulations in arteries, without losing control of the important clinical parameters. The incorporation of cardiovascular autoregulation, together with the well-known impedance boundary condition, forms the basis of the proposed methodology. With autoregulation, the instabilities associated with conventional pressure-type or impedance boundary conditions are avoided without an excessive increase in computational costs. The general behaviour of pulsatile blood flow in arteries, which is important from the clinical point of view, is well reproduced through this new methodology. In addition, the interaction between the blood and the arterial walls occurs via a modified weak coupling, which makes the simulation more stable and computationally efficient. Based on in vitro experiments, the hyperelastic behaviour of the wall is characterised and modelled. The applications and benefits of the proposed pressure-type boundary condition are shown in a model of an idealised aortic arch with and without an ascending aorta dissection, which is a common cardiovascular disorder.

Determinación del riesgo de incendio en una instalación de secado de alperujo = Determination of the risk of fire in an olive-oil mill waste dryer

For the energy valorization of alperujo, residue of the olive oil two phases extraction process, it is necessary to perform a drying process to reduce moisture content from over 60% to less than 10%. In order to reduce primary energy consumption and get an economic return, usually in this kind of drying facilities Gas Turbine CHP is used as a heat source. There have been recently in Spain some fires in this kind of GT-CHP facilities, which have caused high material losses. In some of these fires it has been suggested that the fire was caused by the output of incandescent alperujo in the flue gasesof the drying system. Therefore, the aim of this study is to determine experimentally and analytically under which operational conditions a process of alperujo self-ignition in the drying process can begin, and determine the actual fire hazard in this type of TG-CHP system. For analytical study, the temperature and initial composition of the combustion gases of the Gas Turbine at the entrance of the drying process was calculated and the gas equilibrium conditions reached in contact with the biomass were calculated and, therefore, the temperature of the biomass during the drying process. Moreover, the layer and dust ignition temperature of alperujo has been experimentally determined, according to EN 50281-2-1: 2000. With these results, the operating conditions of the drying process, in which there are real risk of auto-ignition of alperujo have been established.Para la valorización energética del alperujo, residuo del proceso de extracción en dos fases del aceite de oliva, es necesario realizar un proceso de secado para reducir su contenido de humedad de más del 60% al 10% m/m en b.h. Con el fin de reducir el consumo de energía primaria y obtener una rentabilidad económica, normalmente en este tipo de instalaciones de secado se usa la cogeneración con turbina de gas (TG) como fuente de calor. En España en los últimos años han ocurrido algunos casos de incendio en este tipo de instalaciones de cogeneración, que han supuesto pérdidas materiales muy elevadas. Por esta razón, el objetivo de este trabajo es determinar analítica y experimentalmente las condiciones operativas del secadero bajo las cuales podría comenzar un proceso de autoinflamación del alperujo y determinar el riesgo real de incendio en este tipo de instalaciones. Para el estudio analítico, se ha planteado y validado el modelo matemático que permite calcular la temperatura y la composición de los gases de combustión a la entrada y a la salida del secadero, en función de las curvas características de la TG, de las condiciones atmosféricas, del caudal y del grado de humedad de la biomasa tratada. El modelo permite además calcular la temperatura de bulbo húmedo, que es la máxima temperatura que podría alcanzar la biomasa durante el proceso de secado y determinar la cantidad de biomasa que se puede secar completamente en función del caudal y de las condiciones de entrada de los gases de combustión. Con estos resultados y la temperatura mínima de autoinflamación del alperujo determinada experimentalmente siguiendo la norma EN 50281- 2-1:2000, se demuestra que en un proceso de secado de alperujo en condiciones normales de operación no existe riesgo de autoencendido que pueda dar origen a un incendio.

Designing conditions for in vitro formation of amyloid protofilaments and fibrils

We have been able to convert a small α/β protein, acylphosphatase, from its soluble and native form into insoluble amyloid fibrils of the type observed in a range of pathological conditions. This was achieved by allowing slow growth in a solution containing moderate concentrations of trifluoroethanol. When analyzed with electron microscopy, the protein aggregate present in the sample after long incubation times consisted of extended, unbranched filaments of 30–50 Å in width that assemble subsequently into higher order structures. This fibrillar material possesses extensive β-sheet structure as revealed by far-UV CD and IR spectroscopy. Furthermore, the fibrils exhibit Congo red birefringence, increased fluorescence with thioflavine T and cause a red-shift of the Congo red absorption spectrum. All of these characteristics are typical of amyloid fibrils. The results indicate that formation of amyloid occurs when the native fold of a protein is destabilized under conditions in which noncovalent interactions, and in particular hydrogen bonding, within the polypeptide chain remain favorable. We suggest that amyloid formation is not restricted to a small number of protein sequences but is a property common to many, if not all, natural polypeptide chains under appropriate conditions.

Crystal structure of a thermostable type B DNA polymerase from Thermococcus gorgonarius

Most known archaeal DNA polymerases belong to the type B family, which also includes the DNA replication polymerases of eukaryotes, but maintain high fidelity at extreme conditions. We describe here the 2.5 Å resolution crystal structure of a DNA polymerase from the Archaea Thermococcus gorgonarius and identify structural features of the fold and the active site that are likely responsible for its thermostable function. Comparison with the mesophilic B type DNA polymerase gp43 of the bacteriophage RB69 highlights thermophilic adaptations, which include the presence of two disulfide bonds and an enhanced electrostatic complementarity at the DNA–protein interface. In contrast to gp43, several loops in the exonuclease and thumb domains are more closely packed; this apparently blocks primer binding to the exonuclease active site. A physiological role of this “closed” conformation is unknown but may represent a polymerase mode, in contrast to an editing mode with an open exonuclease site. This archaeal B DNA polymerase structure provides a starting point for structure-based design of polymerases or ligands with applications in biotechnology and the development of antiviral or anticancer agents.

Regulation of transport pathways in tumor vessels: Role of tumor type and microenvironment

Novel anti-neoplastic agents such as gene targeting vectors and encapsulated carriers are quite large (approximately 100–300 nm in diameter). An understanding of the functional size and physiological regulation of transvascular pathways is necessary to optimize delivery of these agents. Here we analyze the functional limits of transvascular transport and its modulation by the microenvironment. One human and five murine tumors including mammary and colorectal carcinomas, hepatoma, glioma, and sarcoma were implanted in the dorsal skin-fold chamber or cranial window, and the pore cutoff size, a functional measure of transvascular gap size, was determined. The microenvironment was modulated: (i) spatially, by growing tumors in subcutaneous or cranial locations and (ii) temporally, by inducing vascular regression in hormone-dependent tumors. Tumors grown subcutaneously exhibited a characteristic pore cutoff size ranging from 200 nm to 1.2 μm. This pore cutoff size was reduced in tumors grown in the cranium or in regressing tumors after hormone withdrawal. Vessels induced in basic fibroblast growth factor-containing gels had a pore cutoff size of 200 nm. Albumin permeability was independent of pore cutoff size. These results have three major implications for the delivery of therapeutic agents: (i) delivery may be less efficient in cranial tumors than in subcutaneous tumors, (ii) delivery may be reduced during tumor regression induced by hormonal ablation, and (iii) permeability to a molecule is independent of pore cutoff size as long as the diameter of the molecule is much less than the pore diameter.

Defining the domains of type I collagen involved in heparin- binding and endothelial tube formation

Cell surface heparan sulfate proteoglycan (HSPG) interactions with type I collagen may be a ubiquitous cell adhesion mechanism. However, the HSPG binding sites on type I collagen are unknown. Previously we mapped heparin binding to the vicinity of the type I collagen N terminus by electron microscopy. The present study has identified type I collagen sequences used for heparin binding and endothelial cell–collagen interactions. Using affinity coelectrophoresis, we found heparin to bind as follows: to type I collagen with high affinity (Kd ≈ 150 nM); triple-helical peptides (THPs) including the basic N-terminal sequence α1(I)87–92, KGHRGF, with intermediate affinities (Kd ≈ 2 μM); and THPs including other collagenous sequences, or single-stranded sequences, negligibly (Kd ≫ 10 μM). Thus, heparin–type I collagen binding likely relies on an N-terminal basic triple-helical domain represented once within each monomer, and at multiple sites within fibrils. We next defined the features of type I collagen necessary for angiogenesis in a system in which type I collagen and heparin rapidly induce endothelial tube formation in vitro. When peptides, denatured or monomeric type I collagen, or type V collagen was substituted for type I collagen, no tubes formed. However, when peptides and type I collagen were tested together, only the most heparin-avid THPs inhibited tube formation, likely by influencing cell interactions with collagen–heparin complexes. Thus, induction of endothelial tube morphogenesis by type I collagen may depend upon its triple-helical and fibrillar conformations and on the N-terminal heparin-binding site identified here.

The ability of HIV type 1 to use CCR-3 as a coreceptor is controlled by envelope V1/V2 sequences acting in conjunction with a CCR-5 tropic V3 loop

Although infection by primary HIV type 1 (HIV-1) isolates normally requires the functional interaction of the viral envelope protein with both CD4 and the CCR-5 coreceptor, a subset of such isolates also are able to use the distinct CCR-3 receptor. By analyzing the ability of a series of wild-type and chimeric HIV-1 envelope proteins to mediate CCR-3-dependent infection, we have determined that CCR-3 tropism maps to the V1 and V2 variable region of envelope. Although substitution of the V1/V2 region of a CCR-3 tropic envelope into the context of a CCR-5 tropic envelope is both necessary and sufficient to confer CCR-3 tropism, this same substitution has no phenotypic effect when inserted into a CXCR-4 tropic HIV-1 envelope context. However, this latter chimera acquires both CCR-3 and CCR-5 tropism when a CCR-5 tropic V3 loop sequence also is introduced. These data demonstrate that the V1/2 region of envelope can, like the V3 loop region, encode a particular coreceptor requirement and suggest that a functional envelope:CCR-3 interaction may depend on the cooperative interaction of CCR-3 with both the V1/V2 and the V3 region of envelope.

Antisense oligonucleotides against α1E reduce R-type calcium currents in cerebellar granule cells

Many neurons of the central nervous system display multiple high voltage-activated Ca2+ currents, pharmacologically classified as L-, N-, P-, Q-, and R-type. Of these current types, the R-type is the least understood. The leading candidate for the molecular correlate of R-type currents in cerebellar granule cells is the α1E subunit, which yields Ca2+ currents very similar to the R-type when expressed in heterologous systems. As a complementary approach, we tested whether antisense oligonucleotides against α1E could decrease the expression of R-type current in rat cerebellar granule neurons in culture. Cells were supplemented with either antisense or sense oligonucleotides and whole-cell patch clamp recordings were obtained after 6–8 days in vitro. Incubation with α1E antisense oligonucleotide caused a 52.5% decrease in the peak R-type current density, from −10 ± 0.6 picoamperes/picofarad (pA/pF) (n = 6) in the untreated controls to −4.8 ± 0.8 pA/pF (n = 11) (P < 0.01). In contrast, no significant changes in the current expression were seen in sense oligonucleotide-treated cells (−11.3 ± 3.2 pA/pF). The specificity of the α1E antisense oligonucleotides was supported by the lack of change in estimates of the P/Q current amplitude. Furthermore, antisense and sense oligonucleotides against α1A did not affect R-type current expression (−11.5 ± 1.7 and −11.7 ± 1.7 pA/pF, respectively), whereas the α1A antisense oligonucleotide significantly reduced whole cell currents under conditions in which P/Q current is dominant. Our results support the hypothesis that members of the E class of α1 subunits support the high voltage-activated R-type current in cerebellar granule cells.

Yersinia signals macrophages to undergo apoptosis and YopJ is necessary for this cell death

Pathogenic Yersinia spp. carry a large common plasmid that encodes a number of essential virulence determinants. Included in these factors are the Yersinia-secreted proteins called Yops. We analyzed the consequences of wild-type and mutant strains of Yersinia pseudotuberculosis interactions with the macrophage cell line RAW264.7 and murine bone marrow-derived macrophages. Wild-type Y. pseudotuberculosis kills ≈70% of infected RAW264.7 macrophages and marrow-derived macrophages after an 8-h infection. We show that the cell death mediated by Y. pseudotuberculosis is apoptosis. Mutant Y. pseudotuberculosis that do not make any Yop proteins no longer cause host cell death. Attachment to host cells via invasin or YadA is necessary for the cell death phenotype. Several Yop mutant strains that fail to express one or more Yop proteins were engineered and then characterized for their ability to cause host cell death. A mutant with a polar insertion in YpkA Ser/Thr kinase that does not express YpkA or YopJ is no longer able to cause apoptosis. In contrast, a mutant no longer making YopE or YopH (a tyrosine phosphatase) induces apoptosis in macrophages similar to wild type. When yopJ is added in trans to the ypkAyopJ mutant, the ability of this strain to signal programmed cell death in macrophages is restored. Thus, YopJ is necessary for inducing apoptosis. The ability of Y. pseudotuberculosis to promote apoptosis of macrophages in cell culture suggests that this process is important for the establishment of infection in the host and for evasion of the host immune response.

Copolymer 1 induces T cells of the T helper type 2 that crossreact with myelin basic protein and suppress experimental autoimmune encephalomyelitis

The synthetic amino acid copolymer copolymer 1 (Cop 1) suppresses experimental autoimmune encephalomyelitis (EAE) and is beneficial in multiple sclerosis. To further understand Cop 1 suppressive activity, we studied the cytokine secretion profile of various Cop 1-induced T cell lines and clones. Unlike T cell lines induced by myelin basic protein (MBP), which secreted either T cell helper type 1 (Th1) or both Th1 and Th2 cytokines, the T cell lines/clones induced by Cop 1 showed a progressively polarized development toward the Th2 pathway, until they completely lost the ability to secrete Th1 cytokines. Our findings indicate that the polarization of the Cop 1-induced lines did not result from the immunization vehicle or the in vitro growing conditions, but rather from the tendency of Cop 1 to preferentially induce a Th2 response. The response of all of the Cop 1 specific lines/clones, which were originated in the (SJL/J×BALB/c)F1 hybrids, was restricted to the BALB/c parental haplotype. Even though the Cop 1-induced T cells had not been exposed to the autoantigen MBP, they crossreacted with MBP by secretion of interleukin (IL)-4, IL-6, and IL-10. Administration of these T cells in vivo resulted in suppression of EAE induced by whole mouse spinal cord homogenate, in which several autoantigens may be involved. Secretion of anti-inflammatory cytokines by Cop 1-induced suppressor cells, in response to either Cop 1 or MBP, may explain the therapeutic effect of Cop 1 in EAE and in multiple sclerosis.

Development of an in vitro mRNA decay system for Escherichia coli: Poly(A) polymerase I is necessary to trigger degradation

Using a novel Escherichia coli in vitro decay system in which polysomes are the source of both enzymes and mRNA, we demonstrate a requirement for poly(A) polymerase I (PAP I) in mRNA turnover. The in vitro decay of two different mRNAs (trxA and lpp) is triggered by the addition of ATP only when polysomes are prepared from a strain carrying the wild-type gene for PAP I (pcnB+). The relative decay rates of these two messages are similar in vitro and in vivo. Poly(A) tails are formed on both mRNAs, but no poly(A) tails are detected on the 3′ end of mature 23S rRNA. The size distribution of poly(A) tails generated in vitro, averaging 50 nt in length, is comparable to that previously reported in vivo. PAP I activity is associated exclusively with the polysomes. Exogenously added PAP I does not restore mRNA decay to PAP I− polysomes, suggesting that, in vivo, PAP I may be part of a multiprotein complex. The potential of this in vitro system for analyzing mRNA decay in E. coli is discussed.

Multiple protein domains determine the cell type-specific nuclear distribution of the catalytic subunit required for apolipoprotein B mRNA editing

Apolipoprotein B (apoB) mRNA editing catalyzed by apoB mRNA editing catalytic subunit 1 (APOBEC-1) has been proposed to be a nuclear process. To test this hypothesis, the subcellular distribution of hemagglutinin-(HA) tagged APOBEC-1 expressed in transiently transfected hepatoma cells was determined by indirect immunofluorescence microscopy. HA-APOBEC-1 was detected in both the nucleus and cytoplasm of rat and human hepatoma cells. Mutagenesis of APOBEC-1 demonstrated that the N-terminal 56 amino acids (1–56) were necessary for the nuclear distribution of APOBEC-1, but this region did not contain a functional nuclear localization signal (NLS). However, we identified a 24-amino acid domain in the C terminus of APOBEC-1 with characteristics of a cytoplasmic retention signal (CRS) or a nuclear export signal (NES). These data suggest, therefore, that the nuclear import of APOBEC-1 may not be mediated by a positive NLS; rather, it may be achieved by overcoming the effect of a CRS/NES. We also demonstrated that the nuclear distribution of APOBEC-1 occurred only in cell lines that were capable of editing apoB RNA. We propose that the cellular distribution of APOBEC-1 is determined by multiple domains within this protein, and a nuclear localization of the enzyme may be regulated by cell type-specific factors that render these cells uniquely editing competent.

Transcriptional coupling between the divergent promoters of a prototypic LysR-type regulatory system, the ilvYC operon of Escherichia coli

The twin-domain model [Liu, L. F. & Wang, J. C. (1987) Proc. Natl. Acad. Sci. USA 84, 7024–7027] suggests that closely spaced, divergent, superhelically sensitive promoters can affect the transcriptional activity of one another by transcriptionally induced negative DNA supercoiling generated in the divergent promoter region. This gene arrangement is observed for many LysR-type-regulated operons in bacteria. We have examined the effects of divergent transcription in the prototypic LysR-type system, the ilvYC operon of Escherichia coli. Double-reporter constructs with the lacZ gene under transcriptional control of the ilvC promoter and the galK gene under control of the divergent ilvY promoter were used to demonstrate that a down-promoter mutation in the ilvY promoter severely decreases in vivo transcription from the ilvC promoter. However, a down-promoter mutation in the ilvC promoter only slightly affects transcription from the ilvY promoter. In vitro transcription assays with DNA topoisomers showed that transcription from the ilvC promoter increases over the entire range of physiological superhelical densities, whereas transcription initiation from the ilvY promoter exhibits a broad optimum at a midphysiological superhelical density. Evidence that this promoter coupling is DNA supercoiling-dependent is provided by the observation that a novobiocin-induced decrease in global negative superhelicity results in an increase in ilvY promoter activity and a decrease in ilvC promoter activity predicted by the in vitro data. We suggest that this transcriptional coupling is important for coordinating basal level expression of the ilvYC operon with the nutritional and environmental conditions of cell growth.

Differential use of the signal recognition particle translocase targeting pathway for inner membrane protein assembly in Escherichia coli

Assembly of several inner membrane proteins—leader peptidase (Lep), a Lep derivative (Lep-inv) that inserts with an inverted topology compared with the wild-type protein, the phage M13 procoat protein, and a procoat derivative (H1-procoat) with the hydrophobic core of the signal peptide replaced by a stretch from the first transmembrane segment in Lep—has been studied in vitro and in Escherichia coli strains that are conditional for the expression of either the 54 homologue (Ffh) or 4.5S RNA, which are the two components of the E. coli signal recognition particle (SRP), or SecE, an essential core component of the E. coli preprotein translocase. Membrane insertion has also been tested in a SecB null strain. Lep, Lep-inv, and H1-procoat require SRP for correct assembly into the inner membrane; in contrast, we find that wild-type procoat does not. Lep and, surprisingly, Lep-inv and H1-procoat fail to insert properly when SecE is depleted, whereas insertion of wild-type procoat is unaffected under these conditions. None of the proteins depend on SecB for assembly. These observations indicate that inner membrane proteins can assemble either by a mechanism in which SRP delivers the protein at the preprotein translocase or by what appears to be a direct integration into the lipid bilayer. The observed change in assembly mechanism when the hydrophobicity of the procoat signal peptide is increased demonstrates that the assembly of an inner membrane protein can be rerouted between different pathways.