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首页> 外文期刊>Journal of Molecular Biology >Nhp6p and Med3p regulate gene expression by controlling the local subunit composition of RNA polymerase II.
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Nhp6p and Med3p regulate gene expression by controlling the local subunit composition of RNA polymerase II.

机译：Nhp6p和Med3p通过控制RNA聚合酶II的局部亚基组成来调节基因表达。

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摘要

Nhp6p is an architectural Saccharomyces cerevisiae non-histone chromosomal protein that bends DNA and plays an important role in transcription and genome stability. We used the split-ubiquitin system to isolate proteins that interact with Nhp6p in vivo, and we confirmed 11 of these protein-protein interactions with glutathione S-transferase pull-down experiments in vitro. Most of the Nhp6p-interacting proteins are involved in transcription and DNA repair. We utilized the ZDS1, PUR5 and UME6 genes, which are repressed by Nhp6p and its interacting partners Rpb4p and Med3p, to study the chromosomal localization of these three proteins in wild-type and gene deletion strains. Nhp6p, Med3p and Rpb4p were found at the promoters of ZDS1, PUR5 and UME6, indicating that the repressing effects the three proteins had on the expression of these three genes had been direct ones. We also found that Med3p inhibited promoter clearance of RNA polymerase II, which contained the dissociable subunit Rpb4p, while Nhp6p recruited Rpb4p to the basal promoters of ZDS1, PUR5 and UME6. Our results further suggest that Rpb4p inhibits transcription initiation but stimulates transcription elongation and that Nhp6p and Med3p regulate gene expression by controlling the local subunit composition of RNA polymerase II.
机译：Nhp6p是一种酿酒酵母非组蛋白染色体蛋白质，可弯曲DNA，并在转录和基因组稳定性中起重要作用。我们使用了分裂泛素系统来分离在体内与Nhp6p相互作用的蛋白质，并在体外通过谷胱甘肽S-转移酶下拉实验确认了其中11种蛋白质-蛋白质相互作用。大多数与Nhp6p相互作用的蛋白都参与转录和DNA修复。我们利用被Nhp6p及其相互作用伙伴Rpb4p和Med3p抑制的ZDS1，PUR5和UME6基因，研究了这三种蛋白质在野生型和基因缺失菌株中的染色体定位。在ZDS1，PUR5和UME6的启动子上发现了Nhp6p，Med3p和Rpb4p，这表明这三种蛋白对这三个基因表达的抑制作用是直接的。我们还发现Med3p抑制了RNA聚合酶II的启动子清除，RNA聚合酶II包含可分离的亚基Rpb4p，而Nhp6p则将Rpb4p募集到ZDS1，PUR5和UME6的基础启动子。我们的结果进一步表明，Rpb4p抑制转录启动，但刺激转录延伸，并且Nhp6p和Med3p通过控制RNA聚合酶II的局部亚基组成来调节基因表达。

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