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首页> 外文期刊>Journal of Molecular Biology >ATP binding and hydrolysis by Mcm2 regulate DNA binding by Mcm complexes.
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ATP binding and hydrolysis by Mcm2 regulate DNA binding by Mcm complexes.

机译：ATP与Mcm2的结合和水解可调节Mcm复合物与DNA的结合。

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摘要

The essential minichromosome maintenance (Mcm) proteins Mcm2 through Mcm7 likely comprise the replicative helicase in eukaryotes. In addition to Mcm2-7, other subcomplexes, including one comprising Mcm4, Mcm6, and Mcm7, unwind DNA. Using Mcm4/6/7 as a tool, we reveal a role for nucleotide binding by Saccharomyces cerevisiae Mcm2 in modulating DNA binding by Mcm complexes. Previous studies have shown that Mcm2 inhibits DNA unwinding by Mcm4/6/7. Here, we show that interaction of Mcm2 and Mcm4/6/7 is not sufficient for inhibition; rather, Mcm2 requires nucleotides for its regulatory role. An Mcm2 mutant that is defective for ATP hydrolysis (K549A), as well as ATP analogues, was used to show that ADP binding by Mcm2 is required to inhibit DNA binding and unwinding by Mcm4/6/7. This Mcm2-mediated regulation of Mcm4/6/7 is independent of Mcm3/5. Furthermore, the importance of ATP hydrolysis by Mcm2 to the regulation of the native complex was apparent from the altered DNA binding properties of Mcm2(KA)-7. Moreover, together with the finding that Mcm2(K549A) does not support yeast viability, these results indicate that the nucleotide-bound state of Mcm2 is critical in regulating the activities of Mcm4/6/7 and Mcm2-7 complexes.
机译：基本的微染色体维持（Mcm）蛋白Mcm2至Mcm7可能包含真核生物中的复制解旋酶。除Mcm2-7外，其他子复合物（包括Mcm4，Mcm6和Mcm7的复合物）也展开DNA。使用Mcm4 / 6/7作为工具，我们揭示了酿酒酵母Mcm2核苷酸结合在调节Mcm复合物与DNA结合中的作用。先前的研究表明，Mcm2通过Mcm4 / 6/7抑制DNA解旋。在这里，我们表明Mcm2和Mcm4 / 6/7的相互作用不足以抑制;相反，Mcm2需要核苷酸才能发挥其调节作用。一个有缺陷的ATP水解（M5492）的Mcm2突变体，以及ATP类似物，被用来表明需要Mcm2的ADP结合来抑制DNA的结合和Mcm4 / 6/7的解链。此Mcm2介导的Mcm4 / 6/7调节独立于Mcm3 / 5。此外，从Mcm2（KA）-7改变的DNA结合特性可以明显看出，Mcm2水解ATP对调节天然复合物的重要性。此外，连同发现Mcm2（K549A）不支持酵母活力的发现，这些结果表明Mcm2的核苷酸结合状态对于调节Mcm4 / 6/7和Mcm2-7复合物的活性至关重要。

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