Fares H, Goetsch L, Pringle J R
Department of Biology, University of North Carolina, Chapel Hill 27599, USA.
J Cell Biol. 1996 Feb;132(3):399-411. doi: 10.1083/jcb.132.3.399.
The Saccharomyces cerevisiae CDC3, CDC10, CDC11, and CDC12 genes encode a family of related proteins, the septins, which are involved in cell division and the organization of the cell surface during vegetative growth. A search for additional S. cerevisiae septin genes using the polymerase chain reaction identified SPR3, a gene that had been identified previously on the basis of its sporulation-specific expression. The predicted SPR3 product shows 25-40% identity in amino acid sequence to the previously known septins from S. cerevisiae and other organisms. Immunoblots confirmed the sporulation-specific expression of Spr3p and showed that other septins are also present at substantial levels in sporulating cells. Consistent with the expression data, deletion of SPR3 in either of two genetic backgrounds had no detectable effect on exponentially growing cells. In one genetic background, deletion of SPR3 produced a threefold reduction in sporulation efficiency, although meiosis appeared to be completed normally. In this background, deletion of CDC10 had no detectable effect on sporulation. In the other genetic background tested, the consequences of the two deletions were reversed. Immunofluorescence observations suggest that Spr3p, Cdc3p, and Cdc11p are localized to the leading edges of the membrane sacs that form near the spindle-pole bodies and gradually extend to engulf the nuclear lobes that contain the haploid chromosome sets, thus forming the spores. Deletion of SPR3 does not prevent the localization of Cdc3p and Cdc11p, but these proteins appear to be less well organized, and the intensity of their staining is reduced. Taken together, the results suggest that the septins play important but partially redundant roles during the process of spore formation.
酿酒酵母的CDC3、CDC10、CDC11和CDC12基因编码了一组相关蛋白,即septin蛋白家族,它们在细胞分裂以及营养生长期间细胞表面的组织过程中发挥作用。利用聚合酶链反应寻找酿酒酵母中其他septin基因时,鉴定出了SPR3基因,该基因先前是基于其孢子形成特异性表达而被鉴定出来的。预测的SPR3产物与酿酒酵母及其他生物体中先前已知的septin蛋白在氨基酸序列上有25%-40%的同一性。免疫印迹证实了Spr3p的孢子形成特异性表达,并表明其他septin蛋白在孢子形成细胞中也大量存在。与表达数据一致,在两种遗传背景中的任何一种中缺失SPR3对指数生长的细胞都没有可检测到的影响。在一种遗传背景中,缺失SPR3导致孢子形成效率降低了三倍,尽管减数分裂似乎正常完成。在此背景下,缺失CDC10对孢子形成没有可检测到的影响。在测试的另一种遗传背景中,两种缺失的后果则相反。免疫荧光观察表明,Spr3p、Cdc3p和Cdc11p定位于在纺锤极体附近形成并逐渐延伸以包裹包含单倍体染色体组的核叶的膜囊的前沿,从而形成孢子。缺失SPR3并不妨碍Cdc3p和Cdc11p的定位,但这些蛋白的组织似乎较差,并且它们的染色强度降低。综上所述,结果表明septin蛋白在孢子形成过程中发挥着重要但部分冗余的作用。