Oettinger M A, Struhl K
Mol Cell Biol. 1985 Aug;5(8):1901-9. doi: 10.1128/mcb.5.8.1901-1909.1985.
Transcription of the Saccharomyces cerevisiae his3 gene requires an upstream promoter element and a TATA element. A strain containing his3-delta 13, an allele which deletes the upstream promoter element but contains the TATA box and intact structural gene, fails to express the gene and consequently is unable to grow in medium lacking histidine. In this paper we characterize His+ revertants of his3-delta 13 which are due to unlinked suppressor mutations. Recessive suppressors in three different ope genes allow his3-delta 13 to be expressed at wild-type levels. In all cases, the suppression is due to increased his3 transcription. However, unlike the wild-type his3 gene, whose transcripts are initiated about equally from two different sites (+1 and +12), transcription due to the ope mutations is initiated only from the +12 site, ope-mediated transcription is regulated in a novel manner; it is observed in minimal medium, but not in rich broth. Although ope mutations restore wild-type levels of transcription, his3 chromatin structure, as assayed by micrococcal nuclease sensitivity of the TATA box, resembles that found in the his3-delta 13 parent rather than in the wild-type strain. This provides further evidence that TATA box sensitivity is not correlated with transcriptional activation. ope mutations are pleiotropic in that cells have a crunchy colony morphology and lyse at 37 degrees C in conditions of normal osmolarity. ope mutations are allele specific because they fail to suppress five other his3 promoter mutations. We discuss implications concerning upstream promoter elements and propose some models for ope suppression.
酿酒酵母his3基因的转录需要一个上游启动子元件和一个TATA元件。含有his3-δ13的菌株,该等位基因缺失上游启动子元件,但含有TATA框和完整的结构基因,无法表达该基因,因此不能在缺乏组氨酸的培养基中生长。在本文中,我们对his3-δ13的His+回复突变体进行了表征,这些回复突变体是由非连锁抑制突变引起的。三个不同ope基因中的隐性抑制子使his3-δ13能够以野生型水平表达。在所有情况下,抑制都是由于his3转录增加。然而,与野生型his3基因不同,其转录本大约从两个不同位点(+1和+12)等量起始,ope突变导致的转录仅从+12位点起始,ope介导的转录以一种新的方式受到调控;在基本培养基中可以观察到,但在丰富肉汤中则没有。尽管ope突变恢复了野生型转录水平,但通过TATA框的微球菌核酸酶敏感性测定的his3染色质结构,类似于his3-δ13亲本中的结构,而不是野生型菌株中的结构。这进一步证明TATA框敏感性与转录激活不相关。ope突变具有多效性,因为细胞具有脆的菌落形态,并且在正常渗透压条件下于37℃裂解。ope突变是等位基因特异性的,因为它们不能抑制其他五个his3启动子突变。我们讨论了关于上游启动子元件的意义,并提出了一些ope抑制的模型。