Takegawa Kaoru, Hosomi Akira, Iwaki Tomoko, Fujita Yasuko, Morita Tomotake, Tanaka Naotaka
Department of Life Sciences, Faculty of Agriculture, Kagawa University, Japan.
Biochem Biophys Res Commun. 2003 Nov 7;311(1):77-82. doi: 10.1016/j.bbrc.2003.09.179.
Intracellular vesicle trafficking is mediated by a set of SNARE proteins in eukaryotic cells. Several SNARE proteins are required for vacuolar protein transport and vacuolar biogenesis in Saccharomyces cerevisiae. A search of the Schizosaccharomyces pombe genome database revealed a total of 17 SNARE-related genes. Although no homologs of Vam3p, Nyv1p, and Vam7p have been found in S. pombe, we identified one SNARE-like protein that is homologous to S. cerevisiae Pep12p. However, the disruptants transport vacuolar hydrolase CPY (SpCPY) to the vacuole normally, suggesting that the Pep12 homolog is not required for vacuolar protein transport in S. pombe cells. To identify the SNARE protein(s) involved in Golgi-to-vacuole protein transport, we have deleted four SNARE homolog genes in S. pombe. SpCPY was significantly missorted to the cell surface on deletion of one of the SNARE proteins, Fsv1p (SPAC6F12.03c), with no apparent S. cerevisiae ortholog. In addition, sporulation, endocytosis, and in vivo vacuolar fusion appear to be normal in fsv1Delta cells. These results showed that Fsv1p is mainly involved in vesicle-mediated protein transport between the Golgi and vacuole in S. pombe cells.
真核细胞中的细胞内囊泡运输由一组SNARE蛋白介导。酿酒酵母中的液泡蛋白运输和液泡生物发生需要几种SNARE蛋白。对粟酒裂殖酵母基因组数据库的搜索共发现了17个与SNARE相关的基因。尽管在粟酒裂殖酵母中未发现Vam3p、Nyv1p和Vam7p的同源物,但我们鉴定出了一种与酿酒酵母Pep12p同源的类SNARE蛋白。然而,缺失突变体可将液泡水解酶CPY(SpCPY)正常运输至液泡,这表明粟酒裂殖酵母细胞中的液泡蛋白运输不需要Pep12同源物。为了鉴定参与高尔基体到液泡蛋白运输的SNARE蛋白,我们在粟酒裂殖酵母中删除了四个SNARE同源基因。在缺失一种没有明显酿酒酵母直系同源物的SNARE蛋白Fsv1p(SPAC6F12.03c)后,SpCPY被显著错误分选到细胞表面。此外,fsv1Δ细胞中的孢子形成、内吞作用和体内液泡融合似乎正常。这些结果表明,Fsv1p主要参与粟酒裂殖酵母细胞中高尔基体和液泡之间的囊泡介导的蛋白运输。