FACT facilitates chromatin transcription by RNA polymerases I and III.

FACT facilitates chromatin transcription by RNA polymerases I and III.
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DOI:
10.1038/emboj.2009.33
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发表时间:
2009-04-08
期刊:
影响因子:
11.4
通讯作者:
Zomerdijk, Joost C. B. M.
Zomerdijk, Joost C. B. M.
中科院分区:
生物学1区
文献类型:
--
作者:
Birch, Joanna L.;Tan, Bertrand C-M;Panov, Kostya I.;Panova, Tatiana B.;Andersen, Jens S.;Owen-Hughes, Tom A.;Russell, Jackie;Lee, Sheng-Chung;Zomerdijk, Joost C. B. M.

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从染色质模板的有效转录延伸需要RNA聚合酶(Pos)来协商核小体。我们的生化分析表明,RNA聚合酶I可以通过核小体模板转录,这需要核小体核心颗粒的结构重排。组蛋白伴侣FACT(促进染色质转录)的亚基SSRP 1和Spt 16与哺乳动物Pol I复合物共纯化和共免疫沉淀。在细胞中,SSRP1在rRNA基因重复序列处可检测到。至关重要的是,siRNA介导的FACT亚基表达抑制导致47 S前rRNA水平显着降低,而rRNA前40 nt的合成不受影响,这意味着FACT对于通过染色质延伸Pol I转录很重要。FACT还与RNA Pol III复合物相关,存在于由Pol III转录的基因的染色质中,并促进其在细胞中的转录。我们的研究结果表明,除了在Pol II转录中的既定作用外,FACT在所有三种核RNA Pol的染色质转录中具有生理功能。我们的数据还意味着,当地的染色质动力学影响转录的活性rRNA基因的Pol I和Pol III转录的基因。
Efficient transcription elongation from a chromatin template requires RNA polymerases (Pols) to negotiate nucleosomes. Our biochemical analyses demonstrate that RNA Pol I can transcribe through nucleosome templates and that this requires structural rearrangement of the nucleosomal core particle. The subunits of the histone chaperone FACT (facilitates chromatin transcription), SSRP1 and Spt16, co-purify and co-immunoprecipitate with mammalian Pol I complexes. In cells, SSRP1 is detectable at the rRNA gene repeats. Crucially, siRNA-mediated repression of FACT subunit expression in cells results in a significant reduction in 47S pre-rRNA levels, whereas synthesis of the first 40 nt of the rRNA is not affected, implying that FACT is important for Pol I transcription elongation through chromatin. FACT also associates with RNA Pol III complexes, is present at the chromatin of genes transcribed by Pol III and facilitates their transcription in cells. Our findings indicate that, beyond the established role in Pol II transcription, FACT has physiological functions in chromatin transcription by all three nuclear RNA Pols. Our data also imply that local chromatin dynamics influence transcription of the active rRNA genes by Pol I and of Pol III-transcribed genes.
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