Characterization of the guanine-N7 methyltransferase activity of coronavirus nsp14 on nucleotide GTP.

Characterization of the guanine-N7 methyltransferase activity of coronavirus nsp14 on nucleotide GTP.
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冠状病毒nsp14对核苷酸GTP的鸟嘌呤-N7甲基转移酶活性的表征

DOI:
10.1016/j.virusres.2013.05.001
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发表时间:
2013-09
期刊:
影响因子:
5
通讯作者:
Guo D
Guo D
中科院分区:
医学3区
文献类型:
--
作者:
Jin X;Chen Y;Sun Y;Zeng C;Wang Y;Tao J;Wu A;Yu X;Zhang Z;Tian J;Guo D

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我们发现SARS-CoV nsp 14可以甲基化GTP和dGTP。鉴定了nsp 14对GTP活性所必需的关键残基。m7 GTP或nsp 14可干扰蛋白翻译。大多数在细胞质中复制的真核病毒,包括冠状病毒,都进化出了给它们的RNA加帽的策略。在我们以前的工作中,严重急性呼吸综合征冠状病毒(SARS CoV)的非结构蛋白(nsp)14被鉴定为帽(鸟嘌呤-N7)-甲基转移酶(N7-MTase)。在本研究中,我们发现GTP,dGTP以及帽类似物GpppG,GpppA和m7 GpppG可以被SARS-CoV nsp 14甲基化。相反,nsp 14不能修饰ATP、CTP、UTP、dATP、dCTP、dUTP或帽类似物m7 GpppA。鉴定了nsp 14对GTP上的甲基转移酶活性所必需的关键残基,其包括F73、R84、W86、R310、D331、G333、P335、Y368、C414和C416。我们进一步表明,GTP的甲基转移酶活性对其他冠状病毒的nsp 14是通用的。此外,m7 GTP的积累或蛋白nsp 14的存在可干扰细胞mRNA的蛋白质翻译。总之,结果揭示了冠状病毒nsp 14的一种新的酶活性。
We found SARS-CoV nsp14 could methylate GTP and dGTP. Critical residues of nsp14 essential for the activity on GTP were identified. m7GTP or nsp14 could interfere with protein translation. Most eukaryotic viruses that replicate in the cytoplasm, including coronaviruses, have evolved strategies to cap their RNAs. In our previous work, the nonstructural protein (nsp) 14 of severe acute respiratory syndrome coronavirus (SARS-CoV) was identified as a cap (guanine-N7)-methyltransferase (N7-MTase). In this study, we found that GTP, dGTP as well as cap analogs GpppG, GpppA and m7GpppG could be methylated by SARS-CoV nsp14. In contrast, the nsp14 could not modify ATP, CTP, UTP, dATP, dCTP, dUTP or cap analog m7GpppA. Critical residues of nsp14 essential for the methyltransferase activity on GTP were identified, which include F73, R84, W86, R310, D331, G333, P335, Y368, C414, and C416. We further showed that the methyltransferase activity of GTP was universal for nsp14 of other coronaviruses. Moreover, the accumulation of m7GTP or presence of protein nsp14 could interfere with protein translation of cellular mRNAs. Altogether, the results revealed a new enzymatic activity of coronavirus nsp14.
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