酵母RNA聚合酶II CTD缺失和替换突变体的构建与分析
Construction and analysis of yeast RNA polymerase II CTD deletion and substitution mutations.
作者信息
West M L, Corden J L
机构信息
Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185, USA.
出版信息
Genetics. 1995 Aug;140(4):1223-33. doi: 10.1093/genetics/140.4.1223.
Abstract
The carboxyl-terminal domain (CTD) of the RNA polymerase II largest subunit plays an essential but poorly understood role in transcription. The CTD is highly phosphorylated in vivo and this modification may be important in the transition from transcription initiation to elongation. We report here the development of a strategy for creating novel yeast CTDs. We have used this approach to show that the minimum viable CTD in yeast contains eight consensus (Tyr1Ser2Pro3Thr4Ser5Pro6Ser7) heptapeptide repeats. Substitution of alanine or glutamate for serines in positions two or five is lethal. In addition, changing tyrosine in position one to phenylalanine is lethal. The effects of mutations that alter potential phosphorylation sites are consistent with a requirement for CTD phosphorylation in vivo.
摘要
RNA聚合酶II最大亚基的羧基末端结构域（CTD）在转录过程中起着至关重要但却鲜为人知的作用。CTD在体内高度磷酸化，这种修饰可能在从转录起始到延伸的转变过程中具有重要意义。我们在此报告一种创建新型酵母CTD的策略的开发情况。我们已使用此方法表明酵母中最小的可行CTD包含八个共有（Tyr1Ser2Pro3Thr4Ser5Pro6Ser7）七肽重复序列。将丙氨酸或谷氨酸替代第二或第五位的丝氨酸是致死性的。此外，将第一位的酪氨酸改变为苯丙氨酸也是致死性的。改变潜在磷酸化位点的突变效应与体内CTD磷酸化的需求一致。
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