Komarnitsky P, Cho E J, Buratowski S
Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology, Boston, Massachusetts 02115, USA.
Genes Dev. 2000 Oct 1;14(19):2452-60. doi: 10.1101/gad.824700.
The activities of several mRNA processing factors are coupled to transcription through binding to RNA polymerase II (Pol II). The largest subunit of Pol II contains a repetitive carboxy-terminal domain (CTD) that becomes highly phosphorylated during transcription. mRNA-capping enzyme binds only to phosphorylated CTD, whereas other processing factors may bind to both phosphorylated and unphosphorylated forms. Capping occurs soon after transcription initiation and before other processing events, raising the question of whether capping components remain associated with the transcription complex after they have modified the 5' end of the mRNA. Chromatin immunoprecipitation in Saccharomyces cerevisiae shows that capping enzyme cross-links to promoters but not coding regions. In contrast, the mRNA cap methyltransferase and the Hrp1/CFIB polyadenylation factor cross-link to both promoter and coding regions. Remarkably, the phosphorylation pattern of the CTD changes during transcription. Ser 5 phosphorylation is detected primarily at promoter regions dependent on TFIIH. In contrast, Ser 2 phosphorylation is seen only in coding regions. These results suggest a dynamic association of mRNA processing factors with differently modified forms of the polymerase throughout the transcription cycle.
几种mRNA加工因子的活性通过与RNA聚合酶II(Pol II)结合而与转录偶联。Pol II的最大亚基包含一个重复的羧基末端结构域(CTD),该结构域在转录过程中会发生高度磷酸化。mRNA加帽酶仅与磷酸化的CTD结合,而其他加工因子可能与磷酸化和未磷酸化形式都结合。加帽在转录起始后不久且在其他加工事件之前发生,这就提出了一个问题,即加帽成分在修饰mRNA的5'末端后是否仍与转录复合物相关联。酿酒酵母中的染色质免疫沉淀表明,加帽酶与启动子交联,但不与编码区交联。相反,mRNA帽甲基转移酶和Hrp1/CFIB聚腺苷酸化因子与启动子和编码区都交联。值得注意的是,CTD的磷酸化模式在转录过程中会发生变化。Ser 5磷酸化主要在依赖于TFIIH的启动子区域检测到。相反,Ser 2磷酸化仅在编码区可见。这些结果表明,在整个转录周期中,mRNA加工因子与聚合酶的不同修饰形式存在动态关联。