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Budding Yeast CTDK-I Is Required for DNA Damage-Induced Transcription

机译:DNA损伤诱导转录需要发芽酵母CTDK-I

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CTDK-I phosphorylates the C-terminal domain (CTD) of the large subunit of yeast RNA polymerase II in a reaction that stimulates transcription elongation. Mutations in CTDK-I subunits—Ctk1p, Ctk2p, and Ctk3p—confer conditional phenotypes. In this study, we examined the role of CTDK-I in the DNA damage response. We found that mutation of individual CTDK-I subunits rendered yeast sensitive to hydroxyurea (HU) and UV irradiation. Treatment with DNA-damaging agents increased phosphorylation of Ser2 within the CTD repeats in wild-type but not in ctk1Δ mutant cells. Using microarray hybridization, we identified genes whose transcription following DNA damage is Ctk1p dependent, including several DNA repair and stress response genes. Following HU treatment, the level of Ser2-phosphorylated RNA polymerase II increased both globally and on the CTDK-I-regulated genes. The pleiotropic phenotypes of ctk mutants suggest that CTDK-I activity is essential during large-scale transcriptional repatterning under stress and unfavorable growth conditions.
机译:CTDK-1在刺激转录延伸的反应中磷酸化了酵母RNA聚合酶II大亚基的C末端结构域(CTD)。 CTDK-1亚基(Ctk1p,Ctk2p和Ctk3p)中的突变赋予条件表型。在这项研究中,我们检查了CTDK-1在DNA损伤反应中的作用。我们发现单个CTDK-1亚基的突变使酵母菌对羟基脲(HU)和紫外线辐射敏感。用DNA损伤剂处理可增加野生型CTD重复序列中CTD内Ser2的磷酸化,而 ctk1 Δ突变细胞中则不会。使用微阵列杂交,我们确定了在DNA损伤后其转录是Ctk1p依赖性的基因,包括几个DNA修复和应激反应基因。 HU处理后,Ser2-磷酸化RNA聚合酶II的水平总体上和CTDK-1调控基因上都增加。 ctk 突变体的多效性表型表明,在应激和不利生长条件下,大规模转录重新转录过程中,CTDK-1活性至关重要。

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