The Eps15 C. elegans homologue EHS-1 is implicated in synaptic vesicle recycling

Eps15 represents the prototype of a family of evolutionarily conserved proteins that are characterized by the presence of the EH domain, a protein-protein interaction module, and that are involved in many aspects of intracellular vesicular sorting. Although biochemical and functional studies have implicated Eps15 in endocytosis, its function in the endocytic machinery remains unclear. Here we show that the Caenorhabditis elegans gene, zk1248.3 (ehs-1), is the orthologue of Eps15 in nematodes, and that its product, EHS-1, localizes to synaptic-rich regions. ehs-1-impaired worms showed temperature-dependent depletion of synaptic vesicles and uncoordinated movement. These phenotypes could be correlated with a presynaptic defect in neurotransmission. Impairment of EHS-1 function in dyn-1(ky51) worms, which express a mutant form of dynamin and display a temperature-sensitive locomotion defect, resulted in a worsening of the dyn-1 phenotype and uncoordination at the permissive temperature. Thus, ehs-1 and dyn-1 interact genetically. Moreover, mammalian Eps15 and dynamin protein were shown to interact in vivo. Taken together, our results indicate that EHS-1 acts in synaptic vesicle recycling and that its function might be linked to that of dynamin

Regulation of Fab1 Phosphatidylinositol 3-Phosphate 5-Kinase Pathway by Vac7 Protein and Fig4, a Polyphosphoinositide Phosphatase Family Member

The Saccharomyces cerevisiae FAB1 gene encodes the sole phosphatidylinositol 3-phosphate [PtdIns(3)P] 5-kinase responsible for synthesis of the polyphosphoinositide PtdIns(3,5)P(2). VAC7 encodes a 128-kDa transmembrane protein that localizes to vacuolar membranes. Both vac7 and fab1 null mutants have dramatically enlarged vacuoles and cannot grow at elevated temperatures. Additionally, vac7Δ mutants have nearly undetectable levels of PtdIns(3,5)P(2), suggesting that Vac7 functions to regulate Fab1 kinase activity. To test this hypothesis, we isolated a fab1 mutant allele that bypasses the requirement for Vac7 in PtdIns(3,5)P(2) production. Expression of this fab1 allele in vac7Δ mutant cells suppresses the temperature sensitivity, vacuolar morphology, and PtdIns(3,5)P(2) defects normally exhibited by vac7Δ mutants. We also identified a mutant allele of FIG4, whose gene product contains a Sac1 polyphosphoinositide phosphatase domain, which suppresses vac7Δ mutant phenotypes. Deletion of FIG4 in vac7Δ mutant cells suppresses the temperature sensitivity and vacuolar morphology defects, and dramatically restores PtdIns(3,5)P(2) levels. These results suggest that generation of PtdIns(3,5)P(2) by the Fab1 lipid kinase is regulated by Vac7, whereas turnover of PtdIns(3,5)P(2) is mediated in part by the Sac1 polyphosphoinositide phosphatase family member Fig4

Caracterização morfocultural, biossíntese de autoindutor e formação de biofilme por rizobactérias de hortaliças Morphocultural characterization, autoinducer biosynthesis and biofilm formation in rhizobacteria isolated from vegetable crops

O objetivo deste trabalho foi caracterizar e agrupar rizobactérias, isoladas de hortaliças, quanto à morfologia cultural, riqueza e diversidade e avaliar a biossíntese de autoindutores N-acil lactonas homoserinas (ALH) e a capacidade de formação de biofilmes. Sete estirpes também foram avaliadas quanto ao potencial de promoção de crescimento de Brassica oleraceae var. acephala em casa de vegetação. Para verificar a produção de ALH, foram realizados ensaios com Agrobacterium tumefaciens estirpe NT1 como sistema repórter. A formação de biofilme foi avaliada pelo cultivo do isolado em meio líquido. A promoção do crescimento foi avaliada após inoculação das estirpes em plantas de couve-de-folha pela determinação da produção de massa de matérias fresca e seca. A maior diversidade morfocultural foi encontrada entre as estirpes isoladas de couve-de-folha. De um total de 112 estirpes testadas, 13% foram positivas quanto à produção de ALH; de 91 estirpes, 96% foram capazes de formar biofilmes; e de 79 estirpes, 11% foram positivas para ambas as características. Foram observadas diferenças significativas na massa de matéria seca das raízes com inoculação de 10(9) UFC mL-1 da estirpe R142, que incrementou em 55% a massa de matéria seca das raízes de couve, em relação ao controle. Não há relação entre a capacidade de formar biofilme e a detecção de ALH produzidos pelas rizobactérias avaliadas.<br>The objective of this work was to characterize and group rhizobacteria isolated from vegetable crops for culture morphology, richness and diversity, and to evaluate the biosynthesis of N-acyl homoserine lactone (AHL) autoinducers and the capacity to form biofilms. Seven strains were also assessed for their potential to promote plant growth of Brassica oleraceae var. acephala in greenhouse. To test the production of AHL, the indicator strain Agrobacterium tumefaciens NT1 was used. The formation of biofilms was evaluated by cultivating the isolates in liquid medium. Growth promotion was evaluated after the inoculation of the strains in kale plants and through determination of the fresh and dry mass production. The largest morphocultural diversity was found among the strains isolated from kale. From a total of 112 tested strains, 13% were positive for AHL production, among 91 strains, 96% were capable to form biofilms, and among 79 strains, 11% were positive for both characteristics. Significant differences were observed in root dry mass of plants inoculated with 10(9) UFC mL-1 of strain R142 that increased in 55% the root dry mass in comparison to the control. There is no relationship between the capacity to form biofilms and the detection of the AHL produced by the evaluated rhizobacteria

High-dimensional Exploratory Item Factor Analysis by A Metropolis–Hastings Robbins–Monro Algorithm

stochastic approximation, SA, item response theory, IRT, Markov chain Monte Carlo, MCMC, numerical integration, categorical factor analysis, latent variable modeling, structural equation modeling,