Institute of Molecular and Cell Biology, University of Tartu, Riia 23, Tartu 51010, Estonia.
Mol Cell Biol. 2010 Mar;30(6):1467-77. doi: 10.1128/MCB.01151-09. Epub 2010 Jan 11.
In Saccharomyces cerevisiae SIR proteins mediate transcriptional silencing, forming heterochromatin structures at repressed loci. Although recruitment of transcription initiation factors can occur even to promoters packed in heterochromatin, it is unclear whether heterochromatin inhibits RNA polymerase II (RNAPII) transcript elongation. To clarify this issue, we recruited SIR proteins to the coding region of an inducible gene and characterized the effects of the heterochromatic structure on transcription. Surprisingly, RNAPII is fully competent for transcription initiation and elongation at the locus, leading to significant loss of heterochromatin proteins from the region. A search for auxiliary factors required for transcript elongation through the heterochromatic locus revealed that two proteins involved in histone H3 lysine 56 acetylation, Rtt109 and Asf1, are needed for efficient transcript elongation by RNAPII. The efficiency of transcription through heterochromatin is also impaired in a strain carrying the K56R mutation in histone H3. Our results show that H3 K56 modification is required for efficient transcription of heterochromatic locus by RNAPII, and we propose that transcription-coupled incorporation of H3 acetylated K56 (acK56) into chromatin is needed for efficient opening of heterochromatic loci for transcription.
在酿酒酵母中,SIR 蛋白介导转录沉默,在受抑制的基因座形成异染色质结构。尽管转录起始因子的募集甚至可以发生在包装在异染色质中的启动子上,但异染色质是否抑制 RNA 聚合酶 II(RNAPII)转录延伸尚不清楚。为了澄清这个问题,我们将 SIR 蛋白募集到一个可诱导基因的编码区,并研究了异染色质结构对转录的影响。令人惊讶的是,RNAPII 在该基因座上完全有能力进行转录起始和延伸,导致该区域的异染色质蛋白大量丢失。为了寻找通过异染色质基因座延伸转录所需的辅助因子,我们发现两个参与组蛋白 H3 赖氨酸 56 乙酰化的蛋白质,Rtt109 和 Asf1,对于 RNAPII 的有效转录延伸是必需的。在携带组蛋白 H3 K56 突变的菌株中,通过异染色质的转录效率也受到损害。我们的结果表明,H3 K56 修饰对于 RNAPII 有效转录异染色质基因座是必需的,我们提出转录偶联的组蛋白 H3 乙酰化 K56(acK56)掺入染色质对于异染色质基因座的有效转录开放是必需的。