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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Autoinhibition of Escherichia coli Rep monomer helicase activity by its 2B subdomain
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Autoinhibition of Escherichia coli Rep monomer helicase activity by its 2B subdomain

机译：大肠杆菌Rep单体解旋酶活性的2B亚结构域自动抑制

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摘要

DNA helicases catalyze separation of double-helical DNA into its complementary single strands, a process essential for DNA replication, recombination, and repair. The Escherichia coli Rep protein, a super-family 1 DNA helicase, functions in DNA replication restart and is required for replication of several bacteriophages. Monomers of Rep do not display helicase activity in vitro; in fact, DNA unwinding requires Rep dimerization. Here we show that removal of the 2B subdomain of Rep to form RepΔ2B activates monomer helicase activity, albeit with limited processivity. Although both full length Rep and RepΔ2B monomers can translocate with 3′ to 5′ directionality along single-stranded DNA, the 2B subdomain inhibits the helicase activity of full length Rep. This suggests an autoregulatory mechanism for Rep helicase, which may apply to other nonhexameric helicases, whereby helicase activity is regulated by the rotational conformational state of the 2B subdomain; formation of a Rep dimer may relieve autoinhibition by altering the 2B subdomain orientation.
机译：DNA解旋酶催化双螺旋DNA分离为其互补的单链，这是DNA复制，重组和修复所必需的过程。大肠埃希氏菌Rep蛋白是一种超家族1 DNA解旋酶，可在DNA复制重启中起作用，并且是几种噬菌体复制所必需的。 Rep的单体在体外不显示解旋酶活性。实际上，DNA展开需要Rep二聚化。在这里，我们显示去除Rep的2B子域以形成RepΔ2B可以激活单体解旋酶活性，尽管其合成能力有限。尽管全长Rep和RepΔ2B单体均可沿单链DNA以3'至5'方向易位，但2B亚结构域抑制了全长Rep的解旋酶活性。这表明Rep解旋酶的自调节机制可能适用于其他非六聚体解旋酶，解旋酶的活性受2B子域的旋转构象状态调节； Rep二聚体的形成可以通过改变2B子域方向来缓解自抑制作用。

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