Law D T, Segall J
Department of Biochemistry, University of Toronto, Ontario, Canada.
Mol Cell Biol. 1988 Feb;8(2):912-22. doi: 10.1128/mcb.8.2.912-922.1988.
We previously described the use of a differential hybridization screen of a genomic DNA library of Saccharomyces cerevisiae to identify sporulation-specific (SPS) genes (A. Percival-Smith and J. Segall, Mol. Cell. Biol. 4:142-150, 1984). This initial screen identified 14 SPS genes that are first expressed 6 to 8 h after transfer of cells to sporulation medium. Accumulation of transcripts corresponding to these genes becomes maximal at 8 to 12 h of sporulation, the time at which meiotic events are nearing completion, and by 15 h of sporulation, transcript levels are beginning to decrease. In the present study two additional SPS genes, first expressed at 12 h of sporulation, were isolated. The steady-state level of transcripts corresponding to these two genes, termed SPS100 and SPS101, remains unchanged from 15 to 35 h, a time coincident with spore wall maturation. The nature of the putative 34.2-kilodalton protein encoded by the SPS100 gene is consistent with its being a component of the glycoprotein matrix of the spore wall; the protein contains a potential signal sequence and cleavage site and numerous sites for potential glycosylation. A MATa sps100/MAT alpha sps100 strain was found to be indistinguishable from the wild-type strain when assessed for efficiency of ascus formation and spore viability. However, a more detailed analysis of the mutant strain revealed that the SPS100 gene product serves a protective role during the early stages of spore wall formation. The time at which resistance to ether could first be detected in developing spores was delayed by 5 h in the mutant strain relative to the wild-type strain. This phenotype is presumably a reflection of a defect in spore wall maturation. This study has confirmed that temporally distinct classes of sporulation-specific genes are sequentially activated during the process of meiosis and spore formation and has shown that the SPS100 gene, identified on the basis of its developmental-specific expression pattern, contributes to spore development.
我们之前描述了利用酿酒酵母基因组DNA文库的差异杂交筛选来鉴定孢子形成特异性(SPS)基因(A. Percival-Smith和J. Segall,《分子与细胞生物学》4:142 - 150,1984年)。最初的筛选鉴定出14个SPS基因,这些基因在细胞转移到孢子形成培养基后6至8小时首次表达。对应于这些基因的转录本积累在孢子形成8至12小时达到最大值,此时减数分裂事件接近完成,到孢子形成15小时时,转录本水平开始下降。在本研究中,分离出另外两个在孢子形成12小时首次表达的SPS基因。对应于这两个基因(称为SPS100和SPS101)的转录本稳定状态水平在15至35小时保持不变,这一时期与孢子壁成熟同时发生。由SPS100基因编码的假定34.2千道尔顿蛋白质的性质与其作为孢子壁糖蛋白基质的一个组分相一致;该蛋白质含有一个潜在的信号序列和切割位点以及许多潜在的糖基化位点。当评估子囊形成效率和孢子活力时,发现MATa sps100/MATα sps100菌株与野生型菌株没有区别。然而,对突变菌株更详细的分析表明,SPS100基因产物在孢子壁形成的早期阶段起保护作用。相对于野生型菌株,突变菌株中发育中的孢子首次检测到对乙醚抗性的时间延迟了5小时。这种表型大概反映了孢子壁成熟的缺陷。本研究证实了在减数分裂和孢子形成过程中,不同时间类别的孢子形成特异性基因被依次激活,并且表明基于其发育特异性表达模式鉴定出的SPS100基因对孢子发育有贡献。