Histone H3 lysine 4 methylation is mediated by Set1 and required for cell growth and rDNA silencing in Saccharomyces cerevisiae
Histone H3 lysine 4 methylation is mediated by Set1 and required for cell growth and rDNA silencing in Saccharomyces cerevisiae
复制标题
DOI：
10.1101/gad.940201
复制
发表时间：
2001-12-15
影响因子：
10.5
通讯作者：
Allis, CD
中科院分区：
文献类型：
--
作者：
Briggs, SD;Bryk, M;Allis, CD
已知组蛋白甲基化与转录活性和抑制性染色质状态都相关。最近的研究已经确定了SET结构域的蛋白质，如SUV 39 H1和Clr 4作为介质的H3赖氨酸9（Lys9）甲基化和异染色质的形成。有趣的是，从分离自酿酒酵母或嗜热四膜虫的异步群体的大量组蛋白中未观察到H3 Lys9甲基化。相比之下，H3赖氨酸4（Lys4）甲基化是这些较小真核生物中的主要修饰。为了鉴定负责甲基转移酶并深入了解H3 Lys4甲基化的功能，我们开发了组蛋白H3 Lys4甲基化特异性抗血清。用这种抗血清，我们发现在S.酿酒酵母，结果在体内H3 Lys4甲基化的完全废除。此外，SET1可以挽救set1 Delta菌株中H3 Lys4甲基化的丧失。组蛋白H3在Lys4突变的分析显示了一个缓慢的生长缺陷类似于一个set1三角洲应变。染色质免疫沉淀分析表明，H3 Lys4甲基化存在于rDNA位点，并且Set1介导的H3 Lys4甲基化是rDNA内RNA聚合酶II转录抑制所需的。总之，这些数据表明，Set1介导的H3 Lys4甲基化是正常细胞生长和转录沉默所必需的。
Histone methylation is known to be associated with both transcriptionally active and repressive chromatin states. Recent studies have identified SET domain-containing proteins such as SUV39H1 and Clr4 as mediators of H3 lysine 9 (Lys9) methylation and heterochromatin formation. Interestingly, H3 Lys9 methylation is not observed from bulk histones isolated from asynchronous populations of Saccharomyces cerevisiae or Tetrahymena thermophila. In contrast, H3 lysine 4 (Lys4) methylation is a predominant modification in these smaller eukaryotes. To identify the responsible methyltransferase(s) and to gain insight into the function of H3 Lys4 methylation, we have developed a histone H3 Lys4 methyl-specific antiserum. With this antiserum, we show that deletion of SET1, but not of other putative SET domain-containing genes, in S. cerevisiae, results in the complete abolishment of H3 Lys4 methylation in vivo. Furthermore, loss of H3 Lys4 methylation in a set1 Delta strain can be rescued by SET1. Analysis of histone H3 mutations at Lys4 revealed a slow-growth defect similar to a set1 Delta strain. Chromatin immunoprecipitation assays show that H3 Lys4 methylation is present at the rDNA locus and that Set1-mediated H3 Lys4 methylation is required for repression of RNA polymerase II transcription within rDNA. Taken together, these data suggest that Set1-mediated H3 Lys4 methylation is required for normal cell growth and transcriptional silencing.