Structural basis of nucleosome transcription mediated by Chd1 and FACT.

Structural basis of nucleosome transcription mediated by Chd1 and FACT.
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DOI:
10.1038/s41594-021-00578-6
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发表时间:
2021-04
影响因子:
16.8
通讯作者:
Cramer P
Cramer P
中科院分区:
生物学1区
文献类型:
--
作者:
Farnung L;Ochmann M;Engeholm M;Cramer P

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RNA聚合酶II(Pol II)通过核小体的有效转录需要各种因子的帮助。在这里,我们表明,通过核小体的Pol II转录的生化促进染色质重塑Chd 1和组蛋白伴侣FACT时,延伸因子Spt 4/5和TFIIS存在。我们报告了转录酿酒酵母Pol II− Spt 4/5−核小体复合物与结合Chd 1或FACT的冷冻电镜结构。在第一个结构中,Pol II转录暴露了与Spt 5结合的近端组蛋白H2 A − H2 B二聚体。Pol II还释放了Chd 1的抑制性DNA结合区,该区域准备将DNA泵向Pol II。在第二个结构中,Pol II产生了一个部分解开的核小体,结合FACT,不包括Chd 1和Spt 5。这些结果表明,Pol II通过核小体的进展激活Chd 1,使FACT结合,并最终触发FACT与组蛋白一起转移到上游DNA。RNA聚合酶II-核小体复合物的结构和功能分析揭示了染色质重塑Chd 1和组蛋白伴侣FACT如何通过核小体介导Pol II转录。
Efficient transcription of RNA polymerase II (Pol II) through nucleosomes requires the help of various factors. Here we show biochemically that Pol II transcription through a nucleosome is facilitated by the chromatin remodeler Chd1 and the histone chaperone FACT when the elongation factors Spt4/5 and TFIIS are present. We report cryo-EM structures of transcribing Saccharomyces cerevisiae Pol II−Spt4/5−nucleosome complexes with bound Chd1 or FACT. In the first structure, Pol II transcription exposes the proximal histone H2A−H2B dimer that is bound by Spt5. Pol II has also released the inhibitory DNA-binding region of Chd1 that is poised to pump DNA toward Pol II. In the second structure, Pol II has generated a partially unraveled nucleosome that binds FACT, which excludes Chd1 and Spt5. These results suggest that Pol II progression through a nucleosome activates Chd1, enables FACT binding and eventually triggers transfer of FACT together with histones to upstream DNA. Structural and functional analyses of RNA polymerase II−nucleosome complexes reveal how the chromatin remodeler Chd1 and the histone chaperone FACT mediate Pol II transcription through a nucleosome.
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