Clark-Adams C D, Winston F
Mol Cell Biol. 1987 Feb;7(2):679-86. doi: 10.1128/mcb.7.2.679-686.1987.
Mutations in the Saccharomyces cerevisiae SPT6 gene were originally identified as one class of extragenic suppressors of Ty and delta insertion mutations in the 5' noncoding regions of HIS4 and LYS2. We cloned SPT6 and constructed a null allele by gene disruption. Haploid spores carrying the spt6 null allele were inviable, indicating that the SPT6 gene is essential for mitotic growth. SPT6 was mapped to the right arm of chromosome VII, 44 centimorgans (cM) from ADE6 and 9 cM from CLY8. We showed that spt6 mutations suppress delta insertion mutations at the level of transcription but have no qualitative or quantitative effect on Ty transcription. In addition, we observed interesting SPT6 gene dosage effects. An SPT6 strain containing a high-copy-number plasmid clone of SPT6 showed suppression of delta insertion mutations, and a diploid strain with half its normal dose of SPT6 (SPT6/spt6 null) also exhibited suppression of delta insertion mutations. Therefore, having either too many or too few copies of SPT6 causes a mutant phenotype. Finally, this study and that in the accompanying paper (L. Neigeborn, J. L. Celenza, and M. Carlson, Mol. Cell. Biol. 7:679-686, 1986) showed that spt6 and ssn20 mutations (isolated as suppressors of snf2 and snf5 [sucrose nonfermenting] mutations) identify the same gene. SPT6 and SSN20 have the same genetic map position and share an identical restriction map. Furthermore, spt6 and ssn20 mutations fail to complement each other, and ssn20 mutations suppress solo delta insertion mutations at HIS4 and LYS2. These results, taken in conjunction with the SPT6 dosage effects and the fact that SPT6 is an essential gene, suggest that SPT6 plays a fundamental role in cellular transcription, perhaps by interaction with other transcription factors.
酿酒酵母SPT6基因的突变最初被鉴定为HIS4和LYS2 5'非编码区Ty和δ插入突变的一类基因外抑制子。我们克隆了SPT6并通过基因破坏构建了一个无效等位基因。携带spt6无效等位基因的单倍体孢子无法存活,这表明SPT6基因对有丝分裂生长至关重要。SPT6被定位到染色体VII的右臂,距离ADE6为44厘摩(cM),距离CLY8为9 cM。我们发现spt6突变在转录水平上抑制δ插入突变,但对Ty转录没有定性或定量影响。此外,我们观察到了有趣的SPT6基因剂量效应。含有SPT6高拷贝数质粒克隆的SPT6菌株显示出对δ插入突变的抑制作用,而具有正常剂量一半的SPT6(SPT6/spt6无效)的二倍体菌株也表现出对δ插入突变的抑制作用。因此,SPT6拷贝数过多或过少都会导致突变表型。最后,本研究以及随附论文(L. Neigeborn、J. L. Celenza和M. Carlson,《分子细胞生物学》7:679 - 686,1986)表明,spt6和ssn20突变(作为snf2和snf5 [蔗糖非发酵] 突变的抑制子分离得到)鉴定出同一个基因。SPT6和SSN20具有相同的遗传图谱位置并共享相同的限制性图谱。此外,spt6和ssn20突变不能相互互补,并且ssn20突变抑制HIS4和LYS2处的单个δ插入突变。这些结果,结合SPT6剂量效应以及SPT6是一个必需基因这一事实,表明SPT6可能通过与其他转录因子相互作用在细胞转录中发挥重要作用。