Vba2p, A Vacuolar Membrane Protein Involved in Basic Amino Acid Transport in <i>Schizosaccharomyces pombe</i>

Sugimoto, Naoko; Iwaki, Toru; Chardwiriyapreecha, Soracom; Shimazu, Masamitsu; Sekito, Takayuki; Takegawa, Kaoru; Kakinuma, Yoshihiko

doi:10.1271/bbb.100627

Cited by 6 publications

(6 citation statements)

References 14 publications

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“…The transformed cells were cultured in SD medium without uracil at 30 C until OD 660 reached 1.5, and the vacuolar membranes were stained with lipophilic dye FM4-64 (Invitrogen, Carlsbad, CA), as described previously. 1) As shown in S. pombe cells, 3) GFP-Vba2p was localized exclusively at the vacuolar membrane in S. cerevisiae cells, as shown by co-localization with FM4-64 ( Fig. 1).…”

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confidence: 60%

“…1) In the genome of the fission yeast S. pombe, we have found genes encoding Fnx2p, closest to Fnx1p, 2) and Vba2p, phylogenetically related to the S. cerevisiae VBA family. 3,4) These proteins were localized to the vacuolar membrane of S. pombe cells, and we investigated their roles in amino acid uptake by intact cells. Uptake activities for several amino acids, including lysine, histidine, and arginine, were impaired upon disruption of S. pombe vba2 þ .…”

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confidence: 99%

“…Uptake activities for several amino acids, including lysine, histidine, and arginine, were impaired upon disruption of S. pombe vba2 þ . 3) To characterize these S. pombe transporters biochemically, and more precisely, it is important to establish an in vitro membrane vesicle system, but a method for the purification of vacuoles from S. pombe cells has not been established due to difficulty in the separation of smallsized vacuoles from other organelles such as endosomes. Here we attempted to express the S. pombe vba2 þ gene in S. cerevisiae to evaluate the transport activity of Vba2p using isolated vacuolar membrane vesicles.…”

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Functional Expression ofSchizosaccharomyces pombeVba2p in the Vacuolar Membrane ofSaccharomyces cerevisiae

Pongcharoen

Kawano‐Kawada

Iwaki

et al. 2013

Bioscience, Biotechnology, and Biochemistry

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Functional Expression ofSchizosaccharomyces pombeVba2p in the Vacuolar Membrane ofSaccharomyces cerevisiae

Pongcharoen

Kawano‐Kawada

Iwaki

et al. 2013

Bioscience, Biotechnology, and Biochemistry

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“…Several vacuolar amino acid transporters have been characterized in S. pombe which are homologous to Atg22 (162), Avt3/4 (163,164), Avt5/6 (165,166), and Vba2 (165,167,168) of S. cerevisiae (Fig. 4).…”

Section: Amino Acid Transporters In Other Fungimentioning

confidence: 99%

Regulation of Amino Acid Transport in Saccharomyces cerevisiae

Bianchi

Klooster

Ruiz

et al. 2019

Microbiol Mol Biol Rev

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SUMMARY We review the mechanisms responsible for amino acid homeostasis in Saccharomyces cerevisiae and other fungi. Amino acid homeostasis is essential for cell growth and survival. Hence, the de novo synthesis reactions, metabolic conversions, and transport of amino acids are tightly regulated. Regulation varies from nitrogen pool sensing to control by individual amino acids and takes place at the gene (transcription), protein (posttranslational modification and allostery), and vesicle (trafficking and endocytosis) levels. The pools of amino acids are controlled via import, export, and compartmentalization. In yeast, the majority of the amino acid transporters belong to the APC (amino acid-polyamine-organocation) superfamily, and the proteins couple the uphill transport of amino acids to the electrochemical proton gradient. Although high-resolution structures of yeast amino acid transporters are not available, homology models have been successfully exploited to determine and engineer the catalytic and regulatory functions of the proteins. This has led to a further understanding of the underlying mechanisms of amino acid sensing and subsequent downregulation of transport. Advances in optical microscopy have revealed a new level of regulation of yeast amino acid transporters, which involves membrane domain partitioning. The significance and the interrelationships of the latest discoveries on amino acid homeostasis are put in context.

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“…In addition, Avt5p is involved in the vacuolar uptake of various amino acids [17]. Vba2p is involved in the uptake of basic amino acids into vacuoles [18], and Atg22p is involved in the uptake of several amino acids into vacuoles, as well as in the maintenance of cellular and vacuolar amino acid pools [19]. In any case, establishing an in vitro membrane vesicle system is indispensable for investigating the net transport activities of these transporters.…”

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confidence: 99%

Functional Expression and Characterization of Schizosaccharomyces pombe Avt3p as a Vacuolar Amino Acid Exporter in Saccharomyces cerevisiae

et al. 2015

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In Saccharomyces cerevisiae, Avt3p and Avt4p mediate the extrusion of several amino acids from the vacuolar lumen into the cytosol. SpAvt3p of Schizosaccharomyces pombe, a homologue of these vacuolar amino acid transporters, has been indicated to be involved in spore formation. In this study, we confirmed that GFP-SpAvt3p localized to the vacuolar membrane in S. pombe. The amounts of various amino acids increased significantly in the vacuolar pool of avt3Δ cells, but decreased in that of avt3 +-overexpressing avt3Δ cells. These results suggest that SpAvt3p participates in the vacuolar compartmentalization of amino acids in S. pombe. To examine the export activity of SpAvt3p, we expressed the avt3 + gene in S. cerevisiae cells. We found that the heterologously overproduced GFP-SpAvt3p localized to the vacuolar membrane in S. cerevisiae. Using the vacuolar membrane vesicles isolated from avt3 +-overexpressing S. cerevisiae cells, we detected the export activities of alanine and tyrosine in an ATP-dependent manner. These activities were inhibited by the addition of a V-ATPase inhibitor, concanamycin A, thereby suggesting that the activity of SpAvt3p is dependent on a proton electrochemical gradient generated by the action of V-ATPase. In addition, the amounts of various amino acids in the vacuolar pools of S. cerevisiae cells were decreased by the overproduction of SpAvt3p, which indicated that SpAvt3p was functional in S. cerevisiae cells. Thus, SpAvt3p is a vacuolar transporter that is involved in the export of amino acids from S. pombe vacuoles.

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Vba2p, A Vacuolar Membrane Protein Involved in Basic Amino Acid Transport in Schizosaccharomyces pombe

Cited by 6 publications

References 14 publications

Functional Expression ofSchizosaccharomyces pombeVba2p in the Vacuolar Membrane ofSaccharomyces cerevisiae

Functional Expression ofSchizosaccharomyces pombeVba2p in the Vacuolar Membrane ofSaccharomyces cerevisiae

Regulation of Amino Acid Transport in Saccharomyces cerevisiae

Functional Expression and Characterization of Schizosaccharomyces pombe Avt3p as a Vacuolar Amino Acid Exporter in Saccharomyces cerevisiae

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