Dynamic localization of a cytoplasmic signal transduction response regulator controls morphogenesis during the Caulobacter cell cycle

We present evidence that a bacterial signal transduction cascade that couples morphogenesis with cell cycle progression is regulated by dynamic localization of its components. Previous studies have implicated two histidine kinases, DivJ and PleC, and the response regulator, DivK, in the regulation of morphogenesis in the dimorphic bacterium Caulobacter crescentus. Here, we show that the cytoplasmic response regulator, DivK, exhibits a dynamic, cyclical localization that culminates in asymmetric distribution of DivK within the two cell types that are characteristic of the Caulobacter cell cycle; DivK is dispersed throughout the cytoplasm of the progeny swarmer cell and is localized to the pole of the stalked cell. The membrane-bound DivJ and PleC histidine kinases, which are asymmetrically localized at the opposite poles of the predivisional cell, control the temporal and spatial localization of DivK. DivJ mediates DivK targeting to the poles whereas PleC controls its release from one of the poles at times and places that are consistent with the activities and location of DivJ and PleC in the late predivisional cell. Thus, dynamic changes in subcellular location of multiple components of a signal transduction cascade may constitute a novel mode of prokaryotic regulation to generate and maintain cellular asymmetry.

Multiscale assessment of patterns of avian species richness

The search for a common cause of species richness gradients has spawned more than 100 explanatory hypotheses in just the past two decades. Despite recent conceptual advances, further refinement of the most plausible models has been stifled by the difficulty of compiling high-resolution databases at continental scales. We used a database of the geographic ranges of 2,869 species of birds breeding in South America (nearly a third of the world's living avian species) to explore the influence of climate, quadrat area, ecosystem diversity, and topography on species richness gradients at 10 spatial scales (quadrat area, ≈12,300 to ≈1,225,000 km2). Topography, precipitation, topography × latitude, ecosystem diversity, and cloud cover emerged as the most important predictors of regional variability of species richness in regression models incorporating 16 independent variables, although ranking of variables depended on spatial scale. Direct measures of ambient energy such as mean and maximum temperature were of ancillary importance. Species richness values for 1° × 1° latitude-longitude quadrats in the Andes (peaking at 845 species) were ≈30–250% greater than those recorded at equivalent latitudes in the central Amazon basin. These findings reflect the extraordinary abundance of species associated with humid montane regions at equatorial latitudes and the importance of orography in avian speciation. In a broader context, our data reinforce the hypothesis that terrestrial species richness from the equator to the poles is ultimately governed by a synergism between climate and coarse-scale topographic heterogeneity.

The Schizosaccharomyces pombe spo20+ Gene Encoding a Homologue of Saccharomyces cerevisiae Sec14 Plays an Important Role in Forespore Membrane Formation

The Schizosaccharomyces pombe spo20-KC104 mutation was originally isolated in a screen for sporulation-deficient mutants, and the spo20-KC104 mutant exhibits temperature-sensitive growth. Herein, we report that S. pombe, spo20+ is essential for fission yeast cell viability and is constitutively expressed throughout the life cycle. We also demonstrate that the spo20+ gene product is structurally homologous to Saccharomyces cerevisiae Sec14, the major phosphatidylinositol transfer protein of budding yeast. This structural homology translates to a significant degree of functional relatedness because reciprocal complementation experiments demonstrate that each protein is able to fulfill the essential function of the other. Moreover, biochemical experiments show that, like Sec14, Spo20 is a phosphatidylinositol/phosphatidylcholine-transfer protein. That Spo20 is required for Golgi secretory function in vegetative cells is indicated by our demonstration that the spo20-KC104 mutant accumulates aberrant Golgi cisternae at restrictive temperatures. However, a second phenotype observed in Spo20-deficient fission yeast is arrest of cell division before completion of cell separation. Consistent with a direct role for Spo20 in controlling cell septation in vegetatively growing cells, localization experiments reveal that Spo20 preferentially localizes to the cell poles and to sites of septation of fission yeast cells. We also report that, when fission yeasts are challenged with nitrogen starvation, Spo20 translocates to the nucleus. This nuclear localization persists during conjugation and meiosis. On completion of meiosis, Spo20 translocates to forespore membranes, and it is the assembly of forespore membranes that is abnormal in spo20-KC104 cells. In such mutants, a considerable fraction of forming prespores fail to encapsulate the haploid nucleus. Our results indicate that Spo20 regulates the formation of specialized membrane structures in addition to its recognized role in regulating Golgi secretory function.

Direct observation of extension and retraction of type IV pili

Type IV pili are thin filaments that extend from the poles of a diverse group of bacteria, enabling them to move at speeds of a few tenths of a micrometer per second. They are required for twitching motility, e.g., in Pseudomonas aeruginosa and Neisseria gonorrhoeae, and for social gliding motility in Myxococcus xanthus. Here we report direct observation of extension and retraction of type IV pili in P. aeruginosa. Cells without flagellar filaments were labeled with an amino-specific Cy3 fluorescent dye and were visualized on a quartz slide by total internal reflection microscopy. When pili were attached to a cell and their distal ends were free, they extended or retracted at rates of about 0.5 μm s−1 (29°C). They also flexed by Brownian motion, exhibiting a persistence length of about 5 μm. Frequently, the distal tip of a filament adsorbed to the substratum and the filament was pulled taut. From the absence of lateral deflections of such filaments, we estimate tensions of at least 10 pN. Occasionally, cell bodies came free and were pulled forward by pilus retraction. Thus, type IV pili are linear actuators that extend, attach at their distal tips, exert substantial force, and retract.

Evidence that replication fork components catalyze establishment of cohesion between sister chromatids

Accurate chromosome segregation requires that replicated sister chromatids are held together until anaphase, when their “cohesion” is dissolved, and they are pulled to opposite spindle poles by microtubules. Establishment of new cohesion between sister chromatids in the next cell cycle is coincident with replication fork passage. Emerging evidence suggests that this temporal coupling is not just a coincident timing of independent events, but rather that the establishment of cohesion is likely to involve the active participation of replication-related activities. These include PCNA, a processivity clamp for some DNA polymerases, Trf4/Pol σ (formerly Trf4/Polκ), a novel and essential DNA polymerase, and a modified Replication Factor C clamp–loader complex. Here we describe recent advances in how cohesion establishment is linked to replication, highlight important unanswered questions in this new field, and describe a “polymerase switch” model for how cohesion establishment is coupled to replication fork progression. Building the bridges between newly synthesized sister chromatids appears to be a fundamental but previously unrecognized function of the eukaryotic replication machinery.

Real time observation of anaphase in vitro.

We used digital fluorescence microscopy to make real-time observations of anaphase chromosome movement and changes in microtubule organization in spindles assembled in Xenopus egg extracts. Anaphase chromosome movement in these extracts resembled that seen in living vertebrate cells. During anaphase chromosomes moved toward the spindle poles (anaphase A) and the majority reached positions very close to the spindle poles. The average rate of chromosome to pole movement (2.4 microns/min) was similar to earlier measurements of poleward microtubule flux during metaphase. An increase in pole-to-pole distance (anaphase B) occurred in some spindles. The polyploidy of the spindles we examined allowed us to observe two novel features of mitosis. First, during anaphase, multiple microtubule organizing centers migrated 40 microns or more away from the spindle poles. Second, in telophase, decondensing chromosomes often moved rapidly (7-23 microns/min) away from the spindle poles toward the centers of these asters. This telophase chromosome movement suggests that the surface of decondensing chromosomes, and by extension those of intact nuclei, bear minus-end-directed microtubule motors. Preventing the inactivation of Cdc2/cyclin B complexes by adding nondegradable cyclin B allowed anaphase A to occur at normal velocities, but reduced the ejection of asters from the spindles, blocked chromosome decondensation, and inhibited telophase chromosome movement. In the presence of nondegradable cyclin B, chromosome movement to the poles converted bipolar spindles into pairs of independent monopolar spindles, demonstrating the role of sister chromatid linkage in maintaining spindle bipolarity.

Mos/mitogen-activated protein kinase can induce early meiotic phenotypes in the absence of maturation-promoting factor: a novel system for analyzing spindle formation during meiosis I.

Mitogen-activated protein kinase (MAPK) is selectively activated by injecting either mos or MAPK kinase (mek) RNA into immature mouse oocytes maintained in the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). IBMX arrests oocyte maturation, but Mos (or MEK) overexpression overrides this block. Under these conditions, meiosis I is significantly prolonged, and MAPK becomes fully activated in the absence of p34cdc2 kinase or maturation-promoting factor. In these oocytes, large openings form in the germinal vesicle adjacent to condensing chromatin, and microtubule arrays, which stain for both MAPK and centrosomal proteins, nucleate from these regions. Maturation-promoting factor activation occurs later, concomitant with germinal vesicle breakdown, the contraction of the microtubule arrays into a precursor of the spindle, and the redistribution of the centrosomal proteins into the newly forming spindle poles. These studies define important new functions for the Mos/MAPK cascade in mouse oocyte maturation and, under these conditions, reveal novel detail of the early stages of oocyte meiosis I.

Exchanges are not equally able to enhance meiotic chromosome segregation in yeast.

Homologous chromosomes pair, and then migrate to opposite poles of the spindle at meiosis I. In most eukaryotic organisms, reciprocal recombinations (crossovers) between the homologs are critical to the success of this process. Individuals with defects in meiotic recombination typically produce high levels of aneuploid gametes and exhibit low fertility or are sterile. The experiments described here were designed to test whether different crossovers are equally able to contribute to the fidelity of meiotic chromosome segregation in yeast. These experiments were performed with model chromosomes with which it was possible to control and measure the distributions of meiotic crossovers in wild-type cells. Physical and genetic approaches were used to map crossover positions on model chromosomes and to correlate crossover position with meiotic segregation behavior. The results show that crossovers at different chromosomal positions have different abilities to enhance the fidelity of meiotic segregation.

Proper placement of the Escherichia coli division site requires two functions that are associated with different domains of the MinE protein.

The proper placement of the Escherichia coli division septum requires the MinE protein. MinE accomplishes this by imparting topological specificity to a division inhibitor coded by the minC and minD genes. As a result, the division inhibitor prevents septation at potential division sites that exist at the cell poles but permits septation at the normal division site at midcell. In this paper, we define two functions of MinE that are required for this effect and present evidence that different domains within the 88-amino acid MinE protein are responsible for each of these two functions. The first domain, responsible for the ability of MinE to counteract the activity of the MinCD division inhibitor, is located in a small region near the N terminus of the protein. The second domain, required for the topological specificity of MinE function, is located in the more distal region of the protein and affects the site specificity of placement of the division septum even when separated from the domain responsible for suppression of the activity of the division inhibitor.

Mos overexpression in Swiss 3T3 cells induces meiotic-like alterations of the mitotic spindle.

High levels of mos protooncogene product are expressed during oocyte meiotic maturation and Mos has been implicated in formation of the spindle and spindle pole. Here, we show that in Swiss 3T3 cells with 4N DNA content, high levels of Mos lead to the production of binucleated cells. The Swiss 3T3 cells in mitosis, before binucleation occurs, are anastral and the spindle poles are juxtaposed to the cell membrane. These phenotypes may be related to the meiotic process of attachment of the spindle pole to the oocyte membrane during polar body formation. The production of binucleated somatic cells could result from attachment of the altered mitotic spindle pole to the cell membrane that interferes with cytokinesis but not karyokinesis. This can explain at least one form of genetic instability that leads to altered DNA content in tumor cells.

Notas para una comparación de los proyectos de polos de desarrollo en la Amazonia brasilera y la Patagonia argentina

En este trabajo trazamos líneas de comparación entre los procesos de instalación de planes de desarrollo industrial subsidiados por el Estado nacional en dos grandes regiones de América Latina: la Amazonia brasilera y la Patagonia argentina. Ponemos en debate la noción de desarrollo, enfrentando la igualación que se había construido entre este concepto y el de crecimiento. Con ese objetivo, realizamos una lectura de los planes de desarrollo en su dimensión estructural y nos adentramos en el campo de las luchas sociales y políticas, abarcando para ello un amplio y complejo período histórico

Struggling, Coping, And Thriving: Sense-Making In Stepfamily Couples' Narratives About Coparenting

The term coparenting implies a bioparental dyad that often excludes the stepparent's role in sharing parenting across joint-custody households. Focusing solely on this dyad also precludes gaining an understanding of how stepfamily couples manage together the communication and sharing of parental responsibilities with the parent(s) in the shared children's other home. In a departure from this bioparental dyad-focused approach, this study locates the stepfamily couple at the center of an inquiry into managing coparenting across households. This mixed methods design study included in-depth interviews of 32 stepfamily couples whose narratives about coparenting were analyzed using grounded theory methods. Forty-one percent of stepparents engage in direct coparenting communication, sometimes manifested as the coactive approach identified in this study. Stepfamily couples also involve the stepparent indirectly in coparenting communication, through the conferred and consultative approaches. As well, the couples' narratives about coparenting identify them as either united, where they share the experience, or divided, where coparenting is reserved exclusively for the bioparent to manage. The stepfamily couples' narratives about significant coparenting experiences revealed that they experience and make sense of coparenting as 1) struggling, 2) coping, or 3) thriving. No significant relationship was found between marital satisfaction and experiencing coparenting as strugglers, copers or thrivers. Grounded theory analysis of these narratives also reflects the four dichotomous dimensions of 1) regard-disregard, 2) decency-duplicity, 3) facilitation-interference, and 4) accommodation-inflexibility. Significant incidents located along these dimensions contribute to the stepfamily couples' identification as struggling, coping, or thriving in coparenting. Experiences on the extreme ends of the dichotomous dimensions generate positive and negative turning points for the coparenting interactions and relationships. As well, experiences on the negative end of the dimensional poles can present challenges for the stepfamily couples. Finally, a synthesis of the findings related to the dichotomous dimensions generates a theory of shared parenting values expectancy.

Hybrid convex combinations for IIR system identification.

The low complexity of IIR adaptive filters (AFs) is specially appealing to realtime applications but some drawbacks have been preventing their widespread use so far. For gradient based IIR AFs, adverse operational conditions cause convergence problems in system identification scenarios: underdamped and clustered poles, undermodelling or non-white input signals lead to error surfaces where the adaptation nearly stops on large plateaus or get stuck at sub-optimal local minima that can not be identified as such a priori. Furthermore, the non-stationarity in the input regressor brought by the filter recursivity and the approximations made by the update rules of the stochastic gradient algorithms constrain the learning step size to small values, causing slow convergence. In this work, we propose IIR performance enhancement strategies based on hybrid combinations of AFs that achieve higher convergence rates than ordinary IIR AFs while keeping the stability.

Pion-kaon scattering amplitude constrained with forward dispersion relations up to 1.6 GeV

In this work we provide simple and precise parametrizations of the existing πK scattering data from threshold up to 1.6 GeV, which are constrained to satisfy forward dispersion relations as well as three additional threshold sum rules. We also provide phenomenological values of the threshold parameters and of the resonance poles that appear in elastic scattering.

Cultura organizacional de cursos de ciências contábeis: um estudo em duas universidades públicas

O objetivo geral deste trabalho foi compreender o atual estágio da Cultura Organizacional Acadêmica (COA) em duas Instituições de Ensino Superior (IES) brasileiras, visando identificar possíveis relações entre a cultura e os resultados decorrentes do desempenho dessas IES. A revisão da literatura contemplou as principais teorias sobre cultura organizacional (CO), principalmente as abordagens de Schein (2009), Denison et al. (2012) e Hofstede (2003), além de referenciais empíricos de pesquisadores que se basearam nas abordagens desses autores. A metodologia utilizada teve predomínio qualitativo, com base em etnografia, elementos da observação participante e uso de entrevistas, tanto em profundidade quanto semiestruturadas, que foram gravadas para posterior transcrição e edição. Para o tratamento dos conteúdos das entrevistas, utilizou-se a técnica do Discurso do Sujeito Coletivo (DSC) de Fernando Lefevre e Ana Lefevre (2012), com o emprego do software Qualiquantisoft. O campo empírico abrangeu dois cursos de Ciências Contábeis de duas universidades públicas estaduais, contatadas por acessibilidade. Estes cursos foram o do Departamento de Contabilidade e Atuária da Faculdade de Economia, Administração e Contabilidade da Universidade de São Paulo [FEA/USP] e o do Departamento de Ciências Sociais Aplicadas da Universidade Estadual de Feira de Santana, na Bahia [DCIS/UEFS]. Foram entrevistados 10 docentes do curso da FEA/USP e oito docentes do curso do DCIS/UEFS. Também foi aplicado um questionário para 122 alunos do curso da FEA/USP e 84 alunos do curso do DCIS/UEFS. Os resultados principais são que o atual estágio da COA nos dois cursos das duas IES apresenta-se em polos diametralmente opostos, ou seja, há força na COA do curso da USP, que se reflete na responsabilidade de manter, no presente, os padrões alcançados no passado, e buscar aumentar essa alta referência e visibilidade do curso no país e na América Latina, com orientação estratégica para internacionalização. Em contrapartida, a situação do curso da UEFS é oposta, na qual, de acordo com seus docentes, ele atravessa um período de queda na sua COA, a qual se mantém basicamente adormecida frente às mudanças do ambiente externo, que se reflete em queda dos níveis de qualidade do curso e dos níveis de motivação dos alunos e docentes. Em síntese, os resultados decorrentes do desempenho do curso têm forte dependência da força e solidez de sua COA, materializada na responsabilidade, qualificação e empenho de seus membros (docentes e gestores).