Moonlighting Phosphatase Activity of Klenow DNA Polymerase in the Presence of RNA
Moonlighting Phosphatase Activity of Klenow DNA Polymerase in the Presence of RNA
复制标题
RNA 存在下 Klenow DNA 聚合酶的兼职磷酸酶活性
DOI：
10.1021/acs.biochem.8b00688
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发表时间：
2018
期刊：
影响因子：
2.9
通讯作者：
Zhen Huang
中科院分区：
文献类型：
--
作者：
Qianwei Zhao;Wen Yang;Tong Qin;Zhen Huang
RNA is a key player in the cellular central dogma, including RNA transcription and protein synthesis. However, it is unknown whether RNA can directly interfere with DNA synthesis. Recently, we have foundin vitrothat while binding to DNA polymerase nonspecifically, RNA can transform DNA polymerase to display a moonlighting activity, dNTP phosphatase, in turn interfering with DNA synthesis. This phosphatase activity removes the γ-phosphate from dNTPs (generating dNDPs) and subsequently removes the β-phosphate from the formed dNDPs (generating dNMPs), confirmed by the noncleavable α,β-CH2-dGTP and β,γ-CH2-dGTP analogues. We also found that dGTP is the best substrate for the phosphatase, and the dNTP phosphatase activity is sensitive to the reaction medium. In addition, we have revealed that RNA can tune the activity of closely related proteins and give rise to new catalytic functions with subtle differences. Moreover, we have demonstratedin vitrothat at the lower dNTP level, this phosphatase can directly inhibit DNA synthesis by dNTP depletion, though the phosphatase activity is 690-fold slower than the polymerase activity. Our observationin vitrosuggests a plausible strategy for RNA to directly interfere with DNA polymerase and DNA synthesisin vivo.
