Kurtz S, Shore D
Department of Microbiology, College of Physicians & Surgeons, Columbia University, New York, New York 10032.
Genes Dev. 1991 Apr;5(4):616-28. doi: 10.1101/gad.5.4.616.
RAP1 is a sequence-specific DNA-binding protein essential for cell growth. The occurrence of RAP1-binding sites in many promoter regions, the mating-type gene silencer elements, and telomeres suggests that RAP1 has multiple functions in the cell. To assess its role in transcription, temperature-sensitive mutations in RAP1 were generated. Analysis of rap1ts strains provides evidence that RAP1 functions in both transcriptional activation and silencing of mating-type genes. Several observations indicate that rap1ts strains are defective in the expression of MAT alpha, whose upstream activation sequence (UAS) contains a RAP1-binding site. At nonpermissive temperatures, decreases in MAT alpha steady-state transcript levels can be detected in MAT alpha rap1ts strains. Furthermore, these strains are deficient in alpha-pheromone production and simultaneously express at least two alpha-specific genes. These phenotypes can be reversed by replacing the RAP1-binding site at MAT alpha with a binding site for the GAL4 transcriptional activator. Certain rap1ts alleles have an opposite effect on the silent mating-type locus HMR, which becomes partially derepressed at nonpermissive temperatures.
RAP1是一种对细胞生长至关重要的序列特异性DNA结合蛋白。RAP1结合位点出现在许多启动子区域、交配型基因沉默元件和端粒中,这表明RAP1在细胞中具有多种功能。为了评估其在转录中的作用,生成了RAP1的温度敏感突变体。对rap1ts菌株的分析提供了证据,表明RAP1在交配型基因的转录激活和沉默中均发挥作用。多项观察结果表明,rap1ts菌株在MATα的表达上存在缺陷,其上游激活序列(UAS)包含一个RAP1结合位点。在非允许温度下,可在MATα rap1ts菌株中检测到MATα稳态转录水平的下降。此外,这些菌株在α-信息素产生方面存在缺陷,同时至少表达两个α特异性基因。通过将MATα处的RAP1结合位点替换为GAL4转录激活因子的结合位点,这些表型可以得到逆转。某些rap1ts等位基因对沉默的交配型基因座HMR具有相反的作用,该基因座在非允许温度下会部分去抑制。