Interaction with single-stranded DNA-binding protein localizes ribonuclease HI to DNA replication forks and facilitates R-loop removal.
Interaction with single-stranded DNA-binding protein localizes ribonuclease HI to DNA replication forks and facilitates R-loop removal.
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DOI：
10.1111/mmi.14529
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发表时间：
2020-09
影响因子：
3.6
通讯作者：
Keck JL
中科院分区：
文献类型：
--
作者：
Wolak C;Ma HJ;Soubry N;Sandler SJ;Reyes-Lamothe R;Keck JL
DNA复制复合物（复制体）经常遇到蛋白质和不寻常的核酸结构，这可能会阻碍它们的进展。屏障可以包括当RNA与RNA聚合酶后面的互补DNA模板杂交时形成的转录复合物和R环。细胞编码几种RNA聚合酶和R环清除机制，以限制复制体暴露于这些潜在的障碍物。一种这样的机制是通过核糖核酸酶HI（RNase HI）水解R环。在这里，我们研究了大肠杆菌RNase HI和单链DNA结合蛋白（SSB）之间的相互作用在这个过程中的细胞作用。与SSB的相互作用将RNase HI焦点定位于DNA复制位点。突变rnhA以编码不能与SSB相互作用但保持酶活性的RNA酶HI变体（rnhAK 60 E）消除了RNA酶HI焦点。该突变还在缺乏Rep解旋酶的细胞中产生介质依赖性缓慢生长表型和激活的DNA损伤反应，Rep解旋酶是一种破坏停滞的转录复合物的酶。RNA聚合酶变体被认为增加或减少R环积累，分别增强或抑制rnhAK 60 E rep：：kan菌株的生长表型。这些结果确定了RNase HI/SSB相互作用在帮助清除阻断DNA复制的R环中的细胞作用。DNA复制复合物通常会遇到障碍，如转录复合物和R环，当RNA与RNA聚合酶后面的互补DNA模板杂交时形成。我们发现，RNA酶HI和单链DNA结合蛋白之间的相互作用定位RNA酶HI，以帮助清除阻止DNA复制的R环。
DNA replication complexes (replisomes) routinely encounter proteins and unusual nucleic acid structures that can impede their progress. Barriers can include transcription complexes and R-loops that form when RNA hybridizes with complementary DNA templates behind RNA polymerases. Cells encode several RNA polymerase and R-loop clearance mechanisms to limit replisome exposure to these potential obstructions. One such mechanism is hydrolysis of R-loops by ribonuclease HI (RNase HI). Here, we examine the cellular role of the interaction between Escherichia coli RNase HI and the single-stranded DNA-binding protein (SSB) in this process. Interaction with SSB localizes RNase HI foci to DNA replication sites. Mutation of rnhA to encode an RNase HI variant that cannot interact with SSB but that maintains enzymatic activity (rnhAK60E) eliminates RNase HI foci. The mutation also produces a media-dependent slow-growth phenotype and an activated DNA damage response in cells lacking Rep helicase, which is an enzyme that disrupts stalled transcription complexes. RNA polymerase variants that are thought to increase or decrease R-loop accumulation enhance or suppress, respectively, the growth phenotype of rnhAK60E rep::kan strains. These results identify a cellular role for the RNase HI/SSB interaction in helping to clear R-loops that block DNA replication. DNA replication complexes routinely encounter impediments such as transcription complexes and R-loops that form when RNA hybridizes with complementary DNA templates behind RNA polymerases. We show that interaction between RNase HI and single-stranded DNA-binding protein localizes RNase HI to help clear R-loops that block DNA replication.
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影响因子：
3.6
作者：
Baharoglu Z;Lestini R;Duigou S;Michel B
通讯作者：
Michel B
影响因子：
4.5
作者：
Costes A;Lecointe F;McGovern S;Quevillon-Cheruel S;Polard P
通讯作者：
Polard P
影响因子：
14.9
作者：
Atkinson J;Gupta MK;McGlynn P
通讯作者：
McGlynn P
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
10.5
作者：
Gan, Wenjian;Guan, Zhishuang;Li, Xialu
通讯作者：
Li, Xialu