PRA1 inhibits the extraction of membrane-bound rab GTPase by GDI1

Rab is a family of small Ras-like GTPases regulating intracellular vesicle transport. We have previously reported that prenylated Rab acceptor or PRA1 interacts with Rab GTPases and vesicle-associated membrane protein (VAMP2). Structural prediction programs suggest that PRA1, with its two extensive hydrophobic domains, is likely to be an integral membrane protein. However, subcellular fractionation and immunocytochemical analyses indicated that PRA1 is localized both in the cytosol and tightly associated with the membrane compartment. The membrane-bound form can be partially extracted with physiological buffer and urea, suggesting that PRA1 is an extrinsic membrane protein. Deletion of the carboxyl-terminal domain resulted in a protein that behaved as an integral membrane protein, indicating that this domain plays an essential role in maintaining PRA1 in a soluble state. PRA1 can also bind weakly to GDP dissociation inhibitor (GDI), a protein involved in the solubilization of membrane-bound Rab GTPases. Addition of PRA1 inhibited the extraction of membrane-bound Rab3A by GDI, suggesting that membrane localization of Rab GTPases is dependent on the opposing action of PRA1 and GDI. The binding of Rab and VAMP2 to PRA1 is mutually exclusive such that Rab3A can displace VAMP2 in a preformed VAMP2-PRA1 complex.     

A new species of Ulmeritoides from Brazil (Ephemeroptera: Leptophlebiidae)

The leptophlebiid mayfly genus Ulmeritoides is composed of nine species, three of which have been recorded in Brazil: U. patagianus, U. uruguayensis, and U. misionensis. In the present paper, a new species of this genus, Ulmeritoides oepa sp.nov., is described and illustrated based on fifteen larvae collected in Uraricoera River, state of Roraima, northern Brazil. The species was compared to U. misionensis, U. tifferae, and U. guanacaste, the other species of Ulmeritoides known from the larval stage. U. opea sp.nov. can be distinguished from these species by the following combination of characteristics: 1) medial denticle on anteromedian emargination of labrum much larger than others; 2) anterior tibiae slightly stained black toward the apex; 3) medial femora with few dorsal spines, similar to short thick needles; 4) hind femora with numerous dorsal spines, similar to short thick needles; 5) medial femora with no medial black markings, apical brown markings. The new species seems to be more closely related to U. misionensis, another species in which the anteromedian emargination of the labrum has a median denticle much larger than the others. The studied specimens were deposited in the Invertebrates Collection of the Amazonian Research National Institute, Manaus, state of Amazonas, Brazil, and in the Entomological Collection of the Department of Zoology, Federal University of Rio Janeiro, Rio de Janeiro, state of Rio de Janeiro, Brazil.     

Disruption of Golgi morphology and trafficking in cells expressing mutant prenylated rab acceptor-1

Prenylated Rab acceptor (PRA1) is a protein that binds Rab GTPases and the v-SNARE VAMP2. The protein is localized to the Golgi complex and post-Golgi vesicles. To determine its functional role, we generated a number of point mutations and divided them into three classes based on cellular localization. Class A mutants were retained in the endoplasmic reticulum (ER) and exerted an inhibitory effect on transport of vesicular stomatitis virus envelope glycoprotein (VSVG) from the ER to Golgi as well as to the plasma membrane. Class B mutants exhibited a highly condensed Golgi complex and inhibited exit of anterograde cargo from this organelle. Class C mutants exhibited an intermediate phenotype with Golgi and ER localization along with extensive tubular structures emanating from the Golgi complex. There was a direct correlation between the cellular phenotype and binding to Rab and VAMP2. Class A and C mutants showed a significant decrease in Rab and VAMP2 binding, whereas an increase in binding was observed in the class B mutants. Thus, PRA1 is required for vesicle formation from the Golgi complex and might be involved in recruitment of Rab effectors and SNARE proteins during cargo sequestration.     

Production,characterization and properties of beta-glucosidase and beta-xylosidase from a strain of Aureobasidium sp

beta-glucosidase and beta-xylosidase production by a yeast-like Aureobasidium sp. was carried out during solid-state and submerged fermentation using different carbon sources and crude enzymes were characterized. beta-glucosidase and beta-xylosidase gave optimum activity at pH 2.0-2.5 and 3.0, respectively. These enzymes presented maximal activity at 65 degrees, and were stable in a wide pH range and high temperatures.     

Yeast Diversity Isolated from Grape Musts During Spontaneous Fermentation from a Brazilian Winery

Saccharomyces and non-Saccharomyces yeast species from a winery located in Brazil were identified by ribosomal gene-sequencing analysis. A total of 130 yeast strains were isolated from grape surfaces and musts during alcoholic fermentation from Isabel, Bordeaux, and Cabernet Sauvignon varieties. Samples were submitted to PCR–RFLP analysis and genomic sequencing. Thirteen species were identified: Candida quercitrusa, Candida stellata, Cryptococcus flavescens, Cryptococcus laurentii, Hanseniaspora uvarum, Issatchenkia occidentalis, Issatchenkia orientalis, Issatchenkia terricola, Pichia kluyveri, Pichia guilliermondii, Pichia sp., Saccharomyces cerevisiae, and Sporidiobolus pararoseus. A sequential substitution of species during the different stages of fermentation, with a dominance of non-Saccharomyces yeasts at the beginning, and a successive replacement of species by S. cerevisiae strains at the final steps were observed. This is the first report about the yeast distribution present throughout the alcoholic fermentation in a Brazilian winery, providing supportive information for future studies on their contribution to wine quality.