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一种推定解旋酶(MOT1基因产物)影响基因表达,是酿酒酵母生存所必需的。

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

作者信息

Davis J L, Kunisawa R, Thorner J

机构信息

Department of Molecular and Cell Biology, University of California, Berkeley 94720.

出版信息

Mol Cell Biol. 1992 Apr;12(4):1879-92. doi: 10.1128/mcb.12.4.1879-1892.1992.

Abstract

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.

摘要

将单倍体酵母细胞暴露于交配信息素会诱导一组基因的转录。这种诱导是通过在所有信息素应答基因上游发现的顺式作用DNA序列介导的。尽管STE12基因产物特异性结合此序列元件,并且是基础转录和诱导转录的最大水平所必需的,但并非所有信息素应答基因都以相同的方式受到调控。为了研究是否有其他因素可能在这些基因的转录中起作用,利用遗传筛选来鉴定能够在没有Ste12的情况下组成型表达信息素应答基因的突变体。通过这种方式,我们鉴定出一个隐性单基因突变(mot1,转录调节因子),它增加了几个但不是所有信息素应答基因的基础表达水平。mot1-1等位基因还放宽了对至少另一类上游激活序列的需求,并增强了另一个以前认为不参与交配途径的基因的表达。携带mot1-1的细胞在30℃下生长缓慢,在38℃下无法存活。通过互补这种温度敏感型致死性克隆了MOT1基因。构建无效等位基因证实MOT1是一个必需基因。MOT1定位于第十六号染色体,编码一个1867个氨基酸的大蛋白,该蛋白包含在已知和假定的解旋酶中发现的所有七个保守结构域。MOT1的产物在整个解旋酶区域与酿酒酵母SNF2/SW12和RAD54基因产物具有显著的同源性。

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