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A presumptive helicase (MOT1 gene product) affects gene expression and is required for viability in the yeast Saccharomyces cerevisiae.

机译:推定解旋酶(MOT1基因产物)影响基因表达是酿酒酵母中生存力所必需的。

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摘要

Exposure of a haploid yeast cell to mating pheromone induces transcription of a set of genes. Induction is mediated through a cis-acting DNA sequence found upstream of all pheromone-responsive genes. Although the STE12 gene product binds specifically to this sequence element and is required for maximum levels of both basal and induced transcription, not all pheromone-responsive genes are regulated in an identical manner. To investigate whether additional factors may play a role in transcription of these genes, a genetic screen was used to identify mutants able to express pheromone-responsive genes constitutively in the absence of Ste12. In this way, we identified a recessive, single gene mutation (mot1, for modifier of transcription) which increases the basal level of expression of several, but not all, pheromone-responsive genes. The mot1-1 allele also relaxes the requirement for at least one other class of upstream activating sequence and enhances the expression of another gene not previously thought to be involved in the mating pathway. Cells carrying mot1-1 grow slowly at 30 degrees C and are inviable at 38 degrees C. The MOT1 gene was cloned by complementation of this temperature-sensitive lethality. Construction of a null allele confirmed that MOT1 is an essential gene. MOT1 residues on chromosome XVI and encodes a large protein of 1,867 amino acids which contains all seven of the conserved domains found in known and putative helicases. The product of MOT1 is strikingly homologous to the Saccharomyces cerevisiae SNF2/SW12 and RAD54 gene products over the entire helicase region.
机译:单倍体酵母细胞暴露于交配的信息素会诱导一组基因的转录。诱导是通过在所有信息素应答基因的上游发现的顺式作用DNA序列介导的。尽管STE12基因产物与该序列元件特异性结合,是最大水平的基础转录和诱导转录所必需的,但并非所有信息素应答基因都以相同的方式调节。为了研究其他因素是否可能在这些基因的转录中发挥作用,使用了遗传筛选来鉴定能够在不存在Ste12的情况下组成型表达信息素响应基因的突变体。通过这种方式,我们确定了一个隐性单基因突变(mot1,表示转录的修饰子),该突变增加了一些(但不是全部)信息素反应基因的基础表达水平。 mot1-1等位基因还放宽了对至少一个其他类别的上游激活序列的要求,并增强了以前认为不参与交配途径的另一个基因的表达。携带mot1-1的细胞在30摄氏度时缓慢生长,在38摄氏度时无法存活。通过补充这种对温度敏感的杀伤力,克隆了MOT1基因。无效等位基因的构建证实了MOT1是必需基因。 MOT1残基位于XVI染色体上,编码1867个氨基酸的大蛋白,其中包含在已知和推定解旋酶中发现的所有七个保守结构域。在整个解旋酶区域中,MOT1的产物与酿酒酵母SNF2 / SW12和RAD54基因的产物具有惊人的同源性。

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