Percepción dispersa: Arquitectura y tactilidad en la sociedad de la comunicación

El tema de la tesis se centra en el pensamiento de Walter Benjamin y en el concepto de percepción distraída, desarrollado en el ensayo La obra de arte en la época de su reproducibilidad técnica (1936). El título percepción dispersa deriva de una profundización de los conceptos benjaminianos en términos lingüísticos, adoptando otros significados de la palabra distracción que relacionan este concepto a cuestiones de ámbito espacial y atmosférico. Benjamin sostiene que la arquitectura, en una época en la que se desarrollan y se difunden nuevas técnicas de reproducción/ comunicación, no se puede percibir solo de manera visual sino también táctil, interrelacionando la difusión de nuevas técnicas de reproducción, la tensión entre diferentes modalidades perceptivas y la arquitectura: según él, la dispersión de la vista a favor de la percepción táctil/háptica es un rasgo característico de una sociedad en la que se desarrollan y se difunden nuevas técnicas de reproducción. La tesis profundiza en estos temas, actualizando el concepto de distracción/dispersión para valorar las repercusiones de esta intuición en el ámbito de una parte de la producción arquitectónica contemporánea: se propone una contextualización histórica y una conceptual, relativas a los proyectos recientes de algunos de los autores que profundizan en el impacto de las nuevas tecnologías en el ámbito del proyecto de arquitectura. La hipótesis que se quiere demostrar es que se está desarrollando una tendencia que lleva proyectos o edificios de alto nivel tecnológico y informático desde un ámbito de (supuesto) predominio de la percepción visual, hasta una arquitectura que proporciona experiencias multisensoriales: espacios que se pueden medir con el movimiento, los músculos y el tacto, que desarrollan un conjunto de sensaciones táctiles/hápticas y no solo visuales. Finalmente se quiere verificar si, revertiendo siglos de evolución visual, la visión táctil y cercana siga siendo uno de los polos dialécticos que estructuran la experiencia de la arquitectura, como previsto por Benjamin hace 75 años. ABSTRACT The thesis focuses on the thought of Walter Benjamin and the concept of distracted perception, developed in the essay The Work of Art in the Age of Mechanical Reproduction (1936). The title derives from a linguistic approach to this concept which further explores the meaning of the word distraction, finally relating this kind of perception to spatial and atmospheric issues. Benjamin argues that architecture, thanks to the development of new reproduction/communication technologies, cannot be perceived exclusively in a visual way but also in a tactile way, interrelating new technologies of reproduction, perceptual modalities and architecture: according to him, the dispersion of the view in favor of tactile/ haptic perception is a fundamental characteristic of a technologically developed society. The thesis explores these issues, updating the concept of distraction/dispersion to assess its implications in the works of some architects that are exploring with their buildings the impact of new communication technologies in the field of architectural design. The hypothesis at the base of this work is that we are facing a new trend in this particular field: many of the most advanced buildings provide multisensory experience, and define spaces that can be better perceived with the sense of touch (haptic sense), going beyond a mere visual perception. Finally the thesis wants to verify if, reversing centuries of visual evolution, tactile and close vision remains one of the dialectical poles that stay at the base of the experience of architecture, as foreseen by Benjamin 75 years ago. ABSTRACT La tesi si basa sul pensiero di Walter Benjamin e sul concetto di percezione distratta, sviluppato nel saggio La opera d’arte nell’epoca della sua riproducibilità tecnica (1936). Il titolo, percezione dispersa, deriva da un approfondimento dei concetti benjaminiani in termini linguistici, adottando ulteriori significati della parola che il filosofo utilizza per indicare la distrazione e che relazionano questo termine a questioni di ambito spaziale ed atmosferico. Benjamin sostiene che l’architettura, in un’epoca nella quale si sviluppano e si diffondono nuove tecniche di riproduzione (o meglio: nuove tecnologie di comunicazione), non possa essere percepita solamente in maniera visuale, ma anche tattile, mettendo in relazione la diffusione di nuove tecnologie, la tensione tra differenti modalità percettive e l’architettura: secondo Benjamin, la dispersione della vista a favore di una ricezione tattile/aptica della realtà è una caratteristica tipica di quei momenti storici nei quali si manifestano grandi trasformazioni di ambito sociale e culturale dovute allo sviluppo di nuove tecnologie di comunicazione. La tesi approfondisce questi temi, aggiornando il concetto di distrazione/dispersione per valutarne le ripercussioni su una certa parte della produzione architettonica contemporanea: si propone una contestualizzazione storica ed una concettuale, relative ai progetti di alcuni architetti che lavorano da tempo sull’impatto delle nuove tecnologie nell’ambito del progetto di architettura. La ipotesi che si dimostra con questa tesi è che si sta sviluppando una contro-tendenza tattile, a seguito della quale molti degli edifici tecnologicamente ed informaticamente più avanzati strutturano la relazione con i propri utenti sulla base di esperienze multisensoriali, definendo spazi che possono essere percepiti attraverso una percezione aptica, piuttosto che visuale. In definitiva la tesi verifica che, superando secoli di evoluzione visuale, la visione tattile e ravvicinata e - più in generale - una percezione di tipo aptico continuano ad essere uno dei poli dialettici che strutturano l’esperienza dell’architettura, come previsto da Benjamin più di 75 anni fa.

Josef Albers : la Interacción de la Matière en el arte y en la arquitectura

La materia se presenta ante nosotros en multiplicidad de formas o “apariencias”. A lo largo de la historia se ha reflexionado sobre la relación entre la materia y la forma en distintos campos desde la filosofía hasta la ciencia, pasando por el arte y la arquitectura, moviéndose entre disciplinas desde las más prácticas a las más artísticas, generando posicionamientos opuestos, como el materialismo-idealismo. El concepto de materia a su vez ha ido cambiando a medida que la conciencia humana y la ciencia han ido evolucionando, pasando de considerarse materia como un ente con “masa” a la materia vacía, donde la “masa” es una ilusión que se nos “aparece” dependiendo de la frecuencia con la que vibra su sistema energético. A partir del concepto de “matière” , Josef Albers desarrolla su metodología docente. La matière es más que el aspecto, la “apariencia” que va más allá de la forma cristalizada. Es la forma cambiante que puede adoptar la materia cuando es transformada por el ser humano, dejando su huella sobre ella. Las tres cualidades de la “matière” que el profesor Albers propone en sus ejercicios del Curso Preliminar para desarrollar la “visión” con la “matière” desde la Bauhaus hasta la Universidad de Yale son: Estructural, Factural y Textural. Al desarrollar la observación, teniendo en cuenta estas tres referencias, se descubriá la desvinculación entre lo material y su apariencia desde la honradez. “La discrepancia entre los hechos físicos y el efecto psíquico”. En un proceso constante de ensayo y error se desarrollará la sensibilización individual hacia el material y la evaluación y critica gracias a la dinámica del taller que permite por comparación, aprender y evolucionar como individuo dentro de una sociedad. Esa metodología inductiva regulada por la economía de recursos, promueve el pensamiento creativo, fundamental para producir a través de la articulación un nuevo lenguaje que por medio de la formulación visual exprese nuestra relación con el mundo, con la vida. La vida que constantemente fluye y oscila entre dos polos opuestos, generando interrelaciones que tejen el mundo. Esas interacciones son las que dan vida a la obra artísitica de Albers. PALABRAS CLAVE: materia y matière, estructural factural y textural, vision, hecho físico y efectos psiquico, pensamiento creativo, vida. The matter stands before us in multiple ways or "appearances". Throughout history the relationship between matter and form has been thought out from different fields, from philosophy to science, including art and architecture, moving between disciplines from the most practical to the most artistic generating positions opposites, as materialism-idealism . The concept of matter in turn has changed as the humna consciousness and science have evolved, from being considered as a matter of "mass" to the empty field where the "mass" is an illusion that we "appears" depending on the frequency with which vibrates its energy system. Using the concept of "matière", Josef Albers develops its teaching methodology. The matière is more than the look, the "appearance" that goes beyond the crystallized form. It is the changing form that may take the matter when it is transformed by humans, leaving their mark on it. The three qualities of "matière" that Professor Albers exercises proposed in the Preliminary Course to develop a "vision" with the "matière" from the Bauhaus to Yale are: Structural, Factural and Textural. To develop observation, taking into account these three references, the separation between the material and its appearance was discovered from honesty. "The discrepancy between physical fact and psychic effect." In an ongoing process by trial and error to develop individual sensitzing towards material and critical evaluation through dynamic workshop. The workshop allows for comparison, learn and evolve as an individual within a society. That inductive methodology regulated by the economy of resources, promotes creative thinking, essential to produce through articulation a new language through visual formulation expresses our relationship with the world, with life. Life constantly flowing, oscillates between two opposite poles, creating relationships that weave the world. These interactions are what give life to the artistic work of Albers. KEYWORDS:

Arquitectura modal o ensamblaje de repertorios, procesos y disposiciones

En muchas ocasiones se ha establecido una crítica sobre la excesiva disposicionalidad estética del arquitecto: el abuso subjetivista, un innatismo desligado del entorno socioambiental o su desafección hacia las necesidades y los requerimientos pertinentes del usuario final, buscando, principalmente, la objetivación cultural como expresión personal e individualista. Criticas que responden a un malestar persistente, que subyace a la práctica arquitectónica. En otras ocasiones se han ideado y propuesto tentativas o aproximaciones para la corrección y regulación de dicho “exceso”. En busca de la “pretendida objetividad”, éstas han recurrido a un pasado prístino, a una tradición inconsciente, colectiva e impersonal, a la ausencia de autoría, a lo artesanal y a lo vernáculo. También hacia un futuro tecnocrático: al mesianismo basado en la máquina, la ciencia, los métodos y el progreso tecnológico. La búsqueda de un lenguaje compartido, de un diseño sistemático y de una adecuación funcional han establecido nuevos excesos –repertoriales-, relegando la creatividad y la concepción estética a un plano casi inexistente, subordinándolo a variables de diseño externas a la autoría y la expresión subjetiva del diseñador. Esta tensión se ha saldado en una pugna de binomios categoriales que reducen la realidad a un dualismo aporético y contrapuesto, difícilmente superable. El propósito articulatorio de dichas posturas, en modelos ontológicos integrales, permite la incorporación de dichas categorías bajo estructuras y ensamblajes que despliegan los modos de relación, mediación y producción arquitectónicas. Estos ensamblajes combinan los distintos agentes –disposicionalidades- y sus repertorios en cada una de las fases del proceso arquitectónico, construyendo modos de acción y relación en base a los acoplamientos diferenciales, que devienen en técnicas y dinámicas de producción arquitectónica. Cada modo integrará diversas variables en base a los modelos propuestos -estratos, períodos y polos- y a la elección intencional –“situada”- de cada práctica: la Autonomía Modal Arquitectónica. El producto final de cada modo –o ensamblaje- será inconmensurable, estableciendo sistemas y métodos diversos para la práctica estética, sin que por ello esté garantizado el valor o éxito del resultado. ABSTRACT On many occasions an aesthetic criticism of the excessive disposition of the architect´s subjectivist abuse, innate posture detached from socioenvironmental issues or disaffection regarding the needs and requirements of who -or what- will finally use it has been brought up mainly in pursuit of a cultural objectification as a personal expression and an individualistic one. It is a criticism which goes hand in hand with a persistent uneasiness which is interwoven in the architectural practice. On other occasions we have seen ideas, attempts and approaches developed and proposed in order to correct, improve or adjust that so-called “excess”. In the search for this " expected objectivity" theorists have resorted to delving into a pristine past, an unconscious tradition both collective and personal, to the absence of authorship, regarding craft and vernacular. The pursuit of a common language, a systematic design and a functional adequacy has established new excesses -repertoires-, leaving behind creativity and aesthetic conception at an almost nonexistent level and subordinating it to variables outside the authoring design and subjective expression of the designer. This tension has resulted in a conflict between categorical binomial, which reduces reality to an aporetic and opposed dualism which is not easily overcome. The purpose of such articulatory positions in comprehensive ontological modals allows for the incorporation of those categories under structures and assemblies which display and unfold different models of relationship, mediation and production of architecture. These assemblies combine different agents - disposition - and their repertoires in each of the phases of the architectural process, building actions and relational modes based on the differential couplings, which become techniques and dynamics in architectural production. Each mode will integrate several variables within the proposed ontological models - strata, periods and poles - based on an intentional choice –“situated knowledge”- of each practice: Architectural Modal Autonomy. The final product of each mode -or assembly- will be immeasurable, establishing different systems and methods for aesthetic practice without the value or success of the outcome ever being guaranteed.

La unidad de lo dual : tradición y modernidad en la obra de Tadao Ando y Toyo Ito

La referencia a la tradición como una fuerza capaz de aportar unidad, al abarcar tanto la continuidad como los cambios en las expresiones al margen de la época o las técnicas empleadas, ha sido siempre un componente muy importante de las manifestaciones artísticas de Japón y es la sutil ligazón que las conecta desde el pasado hasta la actualidad. Se entiende aquí que tradición no equivale simplemente a preservación, sino que se trata de una transmisión con una vertiente dual, pues permite una constante evolución sin que se altere su esencia básica. Es de esta forma que la cultura japonesa de la era Edo (1600-1868), con su alto grado de innovación, riqueza y sofisticación, pero además como epílogo histórico previo a la Restauración Meiji de 1868, ha sido la referencia clave en lo relativo a esa mirada a la tradición nipona para el desarrollo de esta tesis. A partir de la segunda mitad del siglo XIX, todas estas características tan genuinas del período Edo quedaron en suspenso y, desde entonces, Japón ha seguido la vía de la modernización (que en muchos aspectos ha sido también la de la occidentalización). Los entrelazamientos de otras dualidades provocados en Japón por los ataques nucleares de 1945 condujeron a una inevitable fusión de aquel mundo físico con el mundo metafísico, de aquella terrible presencia con un tremendo sentimiento de ausencia y, en definitiva, de Oriente con Occidente. El consiguiente impacto sobre la cultura de la nación tuvo una especial repercusión en el ámbito arquitectónico. En este sentido, el pensamiento francés ha desempeñado un papel fundamental en el replanteamiento radical de muchos supuestos esenciales, de muchos conceptos y valores de la cultura occidental, incluyendo los procedentes de la Ilustración, en este mundo contemporáneo que es cada vez más complejo y plural. Éstos y otros hechos otorgan a la cultura japonesa tradicional una cualidad multidimensional (frente a la marcada bidimensionalidad que suele caracterizar a las culturas occidentales) que, tal y como esta tesis doctoral pretende poner de manifiesto, podría revelarse como una «síntesis de contradicciones» en la obra de los arquitectos japoneses Tadao Ando (Osaka, 1941-) y Toyo Ito (Keijo, actual Seúl, 1941-). En el pensamiento oriental una dualidad es entendida como la complementariedad entre dos polos, sólo en apariencia opuestos, que integra dos vertientes de un único concepto. Al igual que la idea de «parejas» budista, concebida según esta doctrina como una unidad entre dos extremos inevitablemente interrelacionados, el objetivo principal sería el de poner de manifiesto cómo la obra de ambos arquitectos trata de resolver los mismos conflictos partiendo de puntos de vista polarizados. ABSTRACT The reference to tradition as a force for unity, encompassing both continuity and changes in expressions regardless of the time or techniques used, has always been a very important component of the artistic manifestations of Japan and it is the subtle link that connects them from the past to the present. It is understood here that tradition does not mean simply preservation, but it is a transmission with a dual aspect, because it allows a constant evolution without altering its basic essence. It is thus that the Japanese culture of the Edo era (1600-1868), with its high degree of innovation, wealth and sophistication, but also as a historical epilogue prior to the Meiji Restoration of 1868 has been the key reference concerning that look to the Japanese tradition for the development of this thesis. From the second half of the 19th century, all these as genuine features of the Edo period remained at a standstill and, since then, Japan has followed the path of modernization (which has also been that of Westernization in many ways). The interlacements of other dualities caused in Japan by the 1945 nuclear attacks led to an inevitable fusion of the physical with the metaphysical world, of that terrible presence with a tremendous sense of absence and, eventually, of East with West. The resulting impact on the culture of the nation had a special repercussion on the architectural field. In this sense, the French thought has played a key role in the radical rethinking of many core assumptions, many concepts and values of Western thought, including those from the Age of Enlightenment, in this contemporary world that is increasingly complex and plural. These and other facts give the traditional Japanese culture a multidimensional quality (opposed to the marked two-dimensionality that usually characterizes Western cultures). As this thesis aims to highlight, this could prove to be a «synthesis of contradictions» in the work of Japanese architects Tadao Ando (Osaka, 1941-) and Toyo Ito (Keijo, current Seoul, 1941-). In the Eastern thought a duality is understood as the complementarity between two poles, only apparently opposite, which integrates two aspects of a single concept. Like the Buddhist notion of «couples» (conceived according to this doctrine as a unity between two inevitably interlinked extremes) the main objective would be to show how the work of both architects tries to resolve the same conflicts starting with polarized viewpoints.

CANTO, ENCANTO E DESENCANTO: um estudo da tensão entre a prática do Canto Coral frente às novas tendências musicais nas Igrejas Batistas na cidade de Campinas.

Nesta pesquisa desenvolvemos um estudo focado na convivência conflituosa entre a música tradicional e a música contemporânea. Foi desenvolvida em um grupo religioso protestante específico, intuindo investigar a hipótese de que há uma tensão entre a prática do Canto Coral e as novas tendências musicais representadas pelas Bandas. Como procedimento metodológico utilizamos uma pesquisa de campo que avaliou a realidade musical em algumas Igrejas Batistas de Campinas. A partir dos resultados obtidos sugerimos ações concretas à práxis pastoral. O primeiro capítulo propõe uma retrospectiva conceitual e histórica do Canto Coral, avaliando sua função antes e depois do processo da reforma e da nova mentalidade musical desenvolvida por Lutero. O segundo capítulo mostra o desenvolvimento da pesquisa realizada junto a seis Igrejas Batistas. Tenta mensurar o gosto musical dos membros dessas comunidades através de um estudo comparativo a partir dos dois pólos: Canto Coral e Banda, ambos, sendo o específico de um campo maior: Tradição e Contemporâneo. O terceiro capítulo, partindo dos impulsos obtidos, desenvolve uma visão crítica sobre os aspectos da tradição, da aceitação à novas tendências, e da tensão propriamente desenvolvida na convivência entre estilos musicais distintos. Mediante tal realidade conflituosa, apresentamos um desafio à práxis pastoral no sentido de se obter possibilidades para uma convivência musical pacífica, considerando a inserção de elementos da contemporaneidade e da tradição na liturgia Batista.

CANTO, ENCANTO E DESENCANTO: um estudo da tensão entre a prática do Canto Coral frente às novas tendências musicais nas Igrejas Batistas na cidade de Campinas.

Nesta pesquisa desenvolvemos um estudo focado na convivência conflituosa entre a música tradicional e a música contemporânea. Foi desenvolvida em um grupo religioso protestante específico, intuindo investigar a hipótese de que há uma tensão entre a prática do Canto Coral e as novas tendências musicais representadas pelas Bandas. Como procedimento metodológico utilizamos uma pesquisa de campo que avaliou a realidade musical em algumas Igrejas Batistas de Campinas. A partir dos resultados obtidos sugerimos ações concretas à práxis pastoral. O primeiro capítulo propõe uma retrospectiva conceitual e histórica do Canto Coral, avaliando sua função antes e depois do processo da reforma e da nova mentalidade musical desenvolvida por Lutero. O segundo capítulo mostra o desenvolvimento da pesquisa realizada junto a seis Igrejas Batistas. Tenta mensurar o gosto musical dos membros dessas comunidades através de um estudo comparativo a partir dos dois pólos: Canto Coral e Banda, ambos, sendo o específico de um campo maior: Tradição e Contemporâneo. O terceiro capítulo, partindo dos impulsos obtidos, desenvolve uma visão crítica sobre os aspectos da tradição, da aceitação à novas tendências, e da tensão propriamente desenvolvida na convivência entre estilos musicais distintos. Mediante tal realidade conflituosa, apresentamos um desafio à práxis pastoral no sentido de se obter possibilidades para uma convivência musical pacífica, considerando a inserção de elementos da contemporaneidade e da tradição na liturgia Batista.

Bipolar localization of Bacillus subtilis topoisomerase IV, an enzyme required for chromosome segregation

In Bacillus subtilis, parE and parC were shown to be essential genes for the segregation of replicated chromosomes. Disruption of either one of these genes resulted in failure of the nucleoid to segregate. Purified ParE and ParC proteins reconstituted to form topoisomerase IV (topo IV), which was highly proficient for ATP-dependent superhelical DNA relaxation and decatenation of interlocked DNA networks. By immunofluorescence microscopy and by directly visualizing fluorescence by using green fluorescence protein fusions, we determined that ParC is localized at the poles of the bacteria in rapidly growing cultures. The bipolar localization of ParC required functional ParE, suggesting that topo IV activity is required for the localization. ParE was found to be distributed uniformly throughout the cell. On the other hand, fluorescence microscopy showed that the GyrA and GyrB subunits of gyrase were associated with the nucleoid. Our results provide a physiologic distinction between DNA gyrase and topo IV. The subcellular localization of topo IV provides physical evidence that it may be part of the bacterial segregation machinery.

Silver-based crystalline nanoparticles, microbially fabricated

One mechanism of silver resistance in microorganisms is accumulation of the metal ions in the cell. Here, we report on the phenomenon of biosynthesis of silver-based single crystals with well-defined compositions and shapes, such as equilateral triangles and hexagons, in Pseudomonas stutzeri AG259. The crystals were up to 200 nm in size and were often located at the cell poles. Transmission electron microscopy, quantitative energy-dispersive x-ray analysis, and electron diffraction established that the crystals comprise at least three different types, found both in whole cells and thin sections. These Ag-containing crystals are embedded in the organic matrix of the bacteria. Their possible potential as organic-metal composites in thin film and surface coating technology is discussed.

The polo-box-dependent induction of ectopic septal structures by a mammalian polo kinase, Plk, in Saccharomyces cerevisiae

Members of the polo subfamily of protein kinases play pivotal roles in cell-cycle control and proliferation. In addition to a high degree of sequence similarity in the kinase domain, polo kinases contain a strikingly conserved motif termed “polo-box” in the noncatalytic C-terminal domain. We have previously shown that the mammalian polo-like kinase Plk is a functional homolog of Saccharomyces cerevisiae Cdc5. Here, we show that, in a polo-box- and kinase activity-dependent manner, ectopic expression of Plk in budding yeast can induce a class of cells with abnormally elongated buds. In addition to localization at spindle poles and cytokinetic neck filaments, Plk induces and localizes to ectopic septin ring structures within the elongated buds. In contrast, mutations in the polo-box abolish both localization to, and induction of, septal structures. Consistent with the polo-box-dependent subcellular localization, the C-terminal domain of Plk, but not its polo-box mutant, is sufficient for subcellular localization. Our data suggest that Plk may contribute a signal to initiate or promote cytokinetic event(s) and that an intact polo-box is required for regulation of these cellular processes.

The MinC component of the division site selection system in Escherichia coli interacts with FtsZ to prevent polymerization

Positioning of the Z ring at the midcell site in Escherichia coli is assured by the min system, which masks polar sites through topological regulation of MinC, an inhibitor of division. To study how MinC inhibits division, we have generated a MalE-MinC fusion that retains full biological activity. We find that MalE-MinC interacts with FtsZ and prevents polymerization without inhibiting FtsZ's GTPase activity. MalE-MinC19 has reduced ability to inhibit division, reduced affinity for FtsZ, and reduced ability to inhibit FtsZ polymerization. These results, along with MinC localization, suggest that MinC rapidly oscillates between the poles of the cell to destabilize FtsZ filaments that have formed before they mature into polar Z rings.

The Yeast Dynactin Complex Is Involved in Partitioning the Mitotic Spindle between Mother and Daughter Cells during Anaphase B

Although vertebrate cytoplasmic dynein can move to the minus ends of microtubules in vitro, its ability to translocate purified vesicles on microtubules depends on the presence of an accessory complex known as dynactin. We have cloned and characterized a novel gene, NIP100, which encodes the yeast homologue of the vertebrate dynactin complex protein p150glued. Like strains lacking the cytoplasmic dynein heavy chain Dyn1p or the centractin homologue Act5p, nip100Δ strains are viable but undergo a significant number of failed mitoses in which the mitotic spindle does not properly partition into the daughter cell. Analysis of spindle dynamics by time-lapse digital microscopy indicates that the precise role of Nip100p during anaphase is to promote the translocation of the partially elongated mitotic spindle through the bud neck. Consistent with the presence of a true dynactin complex in yeast, Nip100p exists in a stable complex with Act5p as well as Jnm1p, another protein required for proper spindle partitioning during anaphase. Moreover, genetic depletion experiments indicate that the binding of Nip100p to Act5p is dependent on the presence of Jnm1p. Finally, we find that a fusion of Nip100p to the green fluorescent protein localizes to the spindle poles throughout the cell cycle. Taken together, these results suggest that the yeast dynactin complex and cytoplasmic dynein together define a physiological pathway that is responsible for spindle translocation late in anaphase.

Katanin Is Responsible for the M-Phase Microtubule-severing Activity in Xenopus Eggs

Microtubules are dynamic structures whose proper rearrangement during the cell cycle is essential for the positioning of membranes during interphase and for chromosome segregation during mitosis. The previous discovery of a cyclin B/cdc2-activated microtubule-severing activity in M-phase Xenopus egg extracts suggested that a microtubule-severing protein might play an important role in cell cycle-dependent changes in microtubule dynamics and organization. However, the isolation of three different microtubule-severing proteins, p56, EF1α, and katanin, has only confused the issue because none of these proteins is directly activated by cyclin B/cdc2. Here we use immunodepletion with antibodies specific for a vertebrate katanin homologue to demonstrate that katanin is responsible for the majority of M-phase severing activity in Xenopus eggs. This result suggests that katanin is responsible for changes in microtubules occurring at mitosis. Immunofluorescence analysis demonstrated that katanin is concentrated at a microtubule-dependent structure at mitotic spindle poles in Xenopus A6 cells and in human fibroblasts, suggesting a specific role in microtubule disassembly at spindle poles. Surprisingly, katanin was also found in adult mouse brain, indicating that katanin may have other functions distinct from its mitotic role.

Yeast Dam1p Is Required to Maintain Spindle Integrity during Mitosis and Interacts with the Mps1p Kinase

We have identified a mutant allele of the DAM1 gene in a screen for mutations that are lethal in combination with the mps1-1 mutation. MPS1 encodes an essential protein kinase that is required for duplication of the spindle pole body and for the spindle assembly checkpoint. Mutations in six different genes were found to be lethal in combination with mps1-1, of which only DAM1 was novel. The remaining genes encode a checkpoint protein, Bub1p, and four chaperone proteins, Sti1p, Hsc82p, Cdc37p, and Ydj1p. DAM1 is an essential gene that encodes a protein recently described as a member of a microtubule binding complex. We report here that cells harboring the dam1-1 mutation fail to maintain spindle integrity during anaphase at the restrictive temperature. Consistent with this phenotype, DAM1 displays genetic interactions with STU1, CIN8, and KAR3, genes encoding proteins involved in spindle function. We have observed that a Dam1p-Myc fusion protein expressed at endogenous levels and localized by immunofluorescence microscopy, appears to be evenly distributed along short mitotic spindles but is found at the spindle poles at later times in mitosis.

Dynamics of Centromeres during Metaphase–Anaphase Transition in Fission Yeast: Dis1 Is Implicated in Force Balance in Metaphase Bipolar Spindle

In higher eukaryotic cells, the spindle forms along with chromosome condensation in mitotic prophase. In metaphase, chromosomes are aligned on the spindle with sister kinetochores facing toward the opposite poles. In anaphase A, sister chromatids separate from each other without spindle extension, whereas spindle elongation takes place during anaphase B. We have critically examined whether such mitotic stages also occur in a lower eukaryote, Schizosaccharomyces pombe. Using the green fluorescent protein tagging technique, early mitotic to late anaphase events were observed in living fission yeast cells. S. pombe has three phases in spindle dynamics, spindle formation (phase 1), constant spindle length (phase 2), and spindle extension (phase 3). Sister centromere separation (anaphase A) rapidly occurred at the end of phase 2. The centromere showed dynamic movements throughout phase 2 as it moved back and forth and was transiently split in two before its separation, suggesting that the centromere was positioned in a bioriented manner toward the poles at metaphase. Microtubule-associating Dis1 was required for the occurrence of constant spindle length and centromere movement in phase 2. Normal transition from phase 2 to 3 needed DNA topoisomerase II and Cut1 but not Cut14. The duration of each phase was highly dependent on temperature.

Chromosomal instability and cytoskeletal defects in oral cancer cells

Oral squamous cell carcinomas are characterized by complex, often near-triploid karyotypes with structural and numerical variations superimposed on the initial clonal chromosomal alterations. We used immunohistochemistry combined with classical cytogenetic analysis and spectral karyotyping to investigate the chromosomal segregation defects in cultured oral squamous cell carcinoma cells. During division, these cells frequently exhibit lagging chromosomes at both metaphase and anaphase, suggesting defects in the mitotic apparatus or kinetochore. Dicentric anaphase chromatin bridges and structurally altered chromosomes with consistent long arms and variable short arms, as well as the presence of gene amplification, suggested the occurrence of breakage–fusion–bridge cycles. Some anaphase bridges were observed to persist into telophase, resulting in chromosomal exclusion from the reforming nucleus and micronucleus formation. Multipolar spindles were found to various degrees in the oral squamous cell carcinoma lines. In the multipolar spindles, the poles demonstrated different levels of chromosomal capture and alignment, indicating functional differences between the poles. Some spindle poles showed premature splitting of centrosomal material, a precursor to full separation of the microtubule organizing centers. These results indicate that some of the chromosomal instability observed within these cancer cells might be the result of cytoskeletal defects and breakage–fusion–bridge cycles.