R7A mutation in N protein renders temperature sensitive phenotype of VSV by affecting its replication and transcription in vitro
R7A mutation in N protein renders temperature sensitive phenotype of VSV by affecting its replication and transcription in vitro
复制标题
N蛋白中的R7A突变通过影响其体外复制和转录而呈现VSV温度敏感表型
DOI：
10.1007/s11262-019-01671-1
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发表时间：
2019-05
期刊：
影响因子：
1.6
通讯作者：
Chen Huimin
中科院分区：
文献类型：
--
作者：
Chen Longyun;Zhou Yanyan;Zhao Min;Chen Huimin
核蛋白(N)包裹的病毒基因组RNA是水疱性口炎病毒(VSV)转录和复制的功能模板。N-RNA复合体的晶体结构表明，RNA紧密隔离在N蛋白的两个叶之间。N-末端臂上的残基(R7)对功能性N-RNA模板的形成非常重要。在本研究中，我们发现在37℃时，单一氨基酸替代(R7A)导致了微小基因组系统中CAT表达的丧失，但在31℃时，R7A对CAT的表达几乎没有影响。进一步的分析表明，R7A只在37℃而不是31℃对VSV的RNA合成和细胞质包涵体的形成有很大的影响。为了进一步研究R7A对病毒复制的影响，我们分别在37℃和31℃下考察了微小基因组系统中NR7A对病毒复制的显性-负效应以及N蛋白R7A突变的重组病毒(RVSVR7A)的单步曲线。我们的结果表明，N末端的R7A突变影响了复制和转录，并诱导了VSV变得对温度敏感。
Viral genomic RNA encapsidated by nucleoprotein (N) forms functional template for the transcription and replication of vesicular stomatitis virus (VSV). The crystal structure of the N-RNA complex shows that RNA is tightly sequestered between the two lobes of the N protein. The residue (R7) in N-terminal arm of N is of great importance to the formation of functional N-RNA template. In our study, we found that single amino acid substitution (R7A) resulted in the loss of CAT expression in vitro minigenome system at 37 °C. But the R7A had little effect on CAT expression at 31 °C. Further analysis showed that R7A had great effects on the RNA synthesis and the formation of cytoplasmic inclusions of VSV only at 37 °C not at 31 °C. For the further investigation of the effect of R7A on virus replication, we checked the dominant-negative effect of NR7Ain minigenome system and the single step curve of recombinant virus with R7A mutation in N protein (rVSVR7A) under 37 °C and 31 °C separately. Our results showed that the mutation of R7A within the N-terminal arm of N affected both replication and transcription and induced VSV to become temperature sensitive.
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影响因子：
5.4
作者：
Tober, Reinhard;Banki, Zoltan;Kimpel, Janine
通讯作者：
Kimpel, Janine
影响因子：
3.7
作者：
T. Das;Bimal K. Chakrabarti;Dhrubajyoti Chattopadhyay;Amiya K. Banerjee
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T. Das;Bimal K. Chakrabarti;Dhrubajyoti Chattopadhyay;Amiya K. Banerjee
影响因子：
5.4
作者：
M. Mellon;S. Emerson
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M. Mellon;S. Emerson
影响因子：
64.8
作者：
Mire CE;Matassov D;Geisbert JB;Latham TE;Agans KN;Xu R;Ota-Setlik A;Egan MA;Fenton KA;Clarke DK;Eldridge JH;Geisbert TW
通讯作者：
Geisbert TW
DOI：
10.1073/pnas.73.5.1504
发表时间：
1976-01-01
影响因子：
11.1
作者：
ABRAHAM, G;BANERJEE, AK
通讯作者：
BANERJEE, AK