Piruat J I, Chávez S, Aguilera A
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Spain.
Genetics. 1997 Dec;147(4):1585-94. doi: 10.1093/genetics/147.4.1585.
We provide genetic evidence that HRS1/PGD1, a yeast gene previously identified as a suppressor of the hyper-recombination phenotype of hpr1, has positive and negative roles in transcriptional regulation. We have analyzed three differently regulated promoters, GAL1, PHO5 and HSP26, by beta-galactosidase assays of lacZ-fused promoters and by Northern analysis of the endogenous genes. Transcription of these promoters was derepressed in hrs1delta mutants under conditions in which it is normally repressed in wild type. Under induced conditions it was either strongly reduced or significantly enhanced depending on the promoter system analyzed. Constitutive transcription was not affected, as determined in ADH1 and TEF2. In addition, Hrs1p was required for mating-factor expression, telomere-linked DNA silencing and DNA supercoiling of plasmids. Furthermore, hrs1delta suppressed Ty-insertion mutations and conferred a Gal- phenotype. Many of these phenotypes also result from mutations in GAL11, SIN4 or RGR1, which encode proteins of the RNA polII mediator. We also show that gal11delta and sin4delta partially suppress the hyper-rec phenotype of hpr1 mutants, although to a lesser extent than hrs1delta. Our results provide new evidence for the connection between hpr1delta-induced deletions and transcription. We discuss the possibility that Hrs1p might be a component of the RNA polII transcription machinery.
我们提供了遗传学证据,证明酵母基因HRS1/PGD1(先前被鉴定为hpr1高重组表型的抑制因子)在转录调控中具有正负双重作用。我们通过对与lacZ融合的启动子进行β-半乳糖苷酶分析以及对内源基因进行Northern分析,研究了三种调控方式不同的启动子GAL1、PHO5和HSP26。在野生型中通常被抑制的条件下,这些启动子在hrs1δ突变体中的转录被解除抑制。在诱导条件下,根据所分析的启动子系统不同,转录要么大幅降低,要么显著增强。如在ADH1和TEF2中所测定的,组成型转录不受影响。此外,Hrs1p是交配因子表达、端粒相关DNA沉默和质粒DNA超螺旋所必需的。此外,hrs1δ抑制Ty插入突变并赋予Gal-表型。许多这些表型也由GAL11、SIN4或RGR1的突变导致,它们编码RNA聚合酶II中介体的蛋白质。我们还表明,gal11δ和sin4δ部分抑制hpr1突变体的高重组表型,尽管程度小于hrs1δ。我们的结果为hpr1δ诱导的缺失与转录之间的联系提供了新证据。我们讨论了Hrs1p可能是RNA聚合酶II转录机制组成部分的可能性。