Snyder M
Department of Biology, Yale University, New Haven, Connecticut 06511.
J Cell Biol. 1989 Apr;108(4):1419-29. doi: 10.1083/jcb.108.4.1419.
A yeast gene, SPA2, was isolated with human anti-spindle pole autoantibodies. The SPA2 gene was fused to the Escherichia coli trpE gene, and polyclonal antibodies were prepared to the fusion protein. Immunofluorescence experiments indicate that the SPA2 gene product has a sharply polarized distribution in yeast cells. In budded cells the SPA2 protein is present at the tip of the bud; in unbudded cells, it is localized to one edge of the cell. When a-cells are induced to form schmoos with alpha-factor, the SPA2 protein is found at the tip of the schmoo. These areas of SPA2 localization correspond to cellular sites expected to be involved in bud formation and/or cell growth. The SPA2 antigen is present in a-cells, alpha-cells, and a/alpha-diploid cells, but is absent in mutant cells in which the SPA2 gene has been disrupted. spa2 mutant cells are viable, but display defects in the direction and control of cell growth. Compared to wild-type cells, spa2 mutant cells have slightly altered budding patterns. Entry into stationary phase is impaired for spa2 mutants, and mutants with one particular allele, spa2-7, form multiple buds under nutrient-limiting conditions. Thus, SPA2 is a newly identified yeast gene that is involved in the direction and control of cell division, and whose gene product localizes to the site of cell growth.
利用人抗纺锤极自身抗体分离出一个酵母基因SPA2。将SPA2基因与大肠杆菌trpE基因融合,并制备针对融合蛋白的多克隆抗体。免疫荧光实验表明,SPA2基因产物在酵母细胞中呈明显的极化分布。在出芽细胞中,SPA2蛋白存在于芽的顶端;在未出芽细胞中,它定位于细胞的一侧边缘。当a细胞被α因子诱导形成接合菌管时,在接合菌管的顶端发现了SPA2蛋白。这些SPA2定位区域对应于预期参与芽形成和/或细胞生长的细胞位点。SPA2抗原存在于a细胞、α细胞和a/α二倍体细胞中,但在SPA2基因已被破坏的突变细胞中不存在。spa2突变细胞是有活力的,但在细胞生长的方向和控制方面表现出缺陷。与野生型细胞相比,spa2突变细胞的出芽模式略有改变。spa2突变体进入稳定期受到损害,具有一个特定等位基因spa2 - 7的突变体在营养限制条件下形成多个芽。因此,SPA2是一个新鉴定的酵母基因,它参与细胞分裂的方向和控制,并且其基因产物定位于细胞生长位点。