The chromatin remodeling factor CHD8 interacts with elongating RNA polymerase II and controls expression of the cyclin E2 gene
The chromatin remodeling factor CHD8 interacts with elongating RNA polymerase II and controls expression of the cyclin E2 gene
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DOI：
10.1093/nar/gkp101
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发表时间：
2009-05-01
影响因子：
14.9
通讯作者：
Reyes, J. C.
中科院分区：
文献类型：
--
作者：
Rodriguez-Paredes, M.;Ceballos-Chavez, M.;Reyes, J. C.
CHD 8是SNF 2家族的染色质重塑ATP酶。我们发现CHD 8的耗尽损害细胞增殖。为了鉴定CHD 8靶基因，我们对CHD 8缺失的细胞进行了转录组学分析，发现CHD 8控制细胞周期蛋白E2（CCNE 2）和胸苷酸合成酶（TYMS）的表达，这两个基因在细胞周期的G1/S转换中表达。CHD 8还能够在瞬时转染实验中共激活CCNE 2启动子。染色质免疫沉淀实验表明，CHD 8直接结合到CCNE 2和TYMS基因的5区。有趣的是，RNA聚合酶II（RNAPII）和CHD 8组成型结合到CCNE 2的5启动子近端区域，而不管细胞周期阶段，因此，CCNE 2的表达。CHD 8的串联染色体结构域在体外特异性结合于在赖氨酸4处二甲基化的组蛋白H3。然而，CHD 8缺失不影响该残基的甲基化水平。我们还表明，CHD 8与RNAPII的延长形式，这是磷酸化的羧基末端结构域（CTD）。此外，CHD 8耗尽的细胞对抑制RNAPII在丝氨酸2处磷酸化的药物过敏，这表明CHD 8是RNAPII转录周期的早期步骤所需的。
CHD8 is a chromatin remodeling ATPase of the SNF2 family. We found that depletion of CHD8 impairs cell proliferation. In order to identify CHD8 target genes, we performed a transcriptomic analysis of CHD8-depleted cells, finding out that CHD8 controls the expression of cyclin E2 (CCNE2) and thymidylate synthetase (TYMS), two genes expressed in the G1/S transition of the cell cycle. CHD8 was also able to co-activate the CCNE2 promoter in transient transfection experiments. Chromatin immunoprecipitation experiments demonstrated that CHD8 binds directly to the 5 region of both CCNE2 and TYMS genes. Interestingly, both RNA polymerase II (RNAPII) and CHD8 bind constitutively to the 5 promoter-proximal region of CCNE2, regardless of the cell-cycle phase and, therefore, of the expression of CCNE2. The tandem chromodomains of CHD8 bind in vitro specifically to histone H3 di-methylated at lysine 4. However, CHD8 depletion does not affect the methylation levels of this residue. We also show that CHD8 associates with the elongating form of RNAPII, which is phosphorylated in its carboxy-terminal domain (CTD). Furthermore, CHD8-depleted cells are hypersensitive to drugs that inhibit RNAPII phosphorylation at serine 2, suggesting that CHD8 is required for an early step of the RNAPII transcription cycle.