Mutational analysis of the functional sites in porcine reproductive and respiratory syndrome virus non-structural protein 10.
Mutational analysis of the functional sites in porcine reproductive and respiratory syndrome virus non-structural protein 10.
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猪繁殖与呼吸综合征病毒非结构蛋白10功能位点的突变分析。
DOI：
10.1099/jgv.0.000004
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发表时间：
2015-03
影响因子：
3.8
通讯作者：
Song Yunfeng
中科院分区：
文献类型：
--
作者：
Zhang Yumeng;Li Huan;Peng Guiqing;Zhang Yong;Gao Xiao;Xiao Shaobo;Cao Shengbo;Chen Huanchun;Song Yunfeng
猪繁殖与呼吸综合征病毒（PRRSV）在世界各地流行，给养猪业造成了重大的经济损失。动脉病毒非结构蛋白10 （nsp10）是参与病毒复制多个过程的超家族1解旋酶。然而，PRRSV nsp10尚未得到很好的鉴定。本研究构建了一系列nsp10突变体，并分析了不同酶活性的功能位点。我们发现nsp10可以结合ssDNA和dsDNA，并且这种结合活性可以被Cys25和His32的突变灭活。这两个突变在不影响atp酶活性的情况下也消除了解绕活性。此外，用Ser取代Ala227消除了解旋酶活性，而用Val取代Ala227则增强了解旋酶活性。综上所述，我们的研究结果表明，PRRSV nsp10中的Cys25和His32对核酸结合和解绕至关重要，而Ala227在解旋酶活性中起重要作用。
Porcine reproductive and respiratory syndrome virus (PRRSV) is prevalent throughout the world and has caused major economic losses to the pig industry. Arterivirus non-structural protein 10 (nsp10) is a superfamily 1 helicase participating in multiple processes of virus replication. PRRSV nsp10, however, has not yet been well characterized. In this study, a series of nsp10 mutants were constructed and analysed for functional sites of different enzymic activities. We found that nsp10 could bind both ssDNA and dsDNA, and this binding activity could be inactivated by mutations at Cys25 and His32. These two mutations also abolished unwinding activity without affecting ATPase activity. In addition, substitution of Ala227 by Ser eliminated helicase activity, whilst substitution by Val enhanced unwinding activity. Taken together, our results showed that Cys25 and His32 in PRRSV nsp10 were critical for nucleic acid binding and unwinding, and that Ala227 played an important role in helicase activity.
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影响因子：
6.7
作者：
The PLOS Pathogens Staff
通讯作者：
The PLOS Pathogens Staff
影响因子：
5
作者：
Fang Y;Snijder EJ
通讯作者：
Snijder EJ
影响因子：
1.6
作者：
Li, Yan;Xue, Chunyi;Cao, Yongchang
通讯作者：
Cao, Yongchang
DOI：
10.2741/s367
发表时间：
2013
期刊：
Frontiers in bioscience
影响因子：
--
作者：
Neville S. Gilhooly;Emma J. Gwynn;M. Dillingham
通讯作者：
Neville S. Gilhooly;Emma J. Gwynn;M. Dillingham
影响因子：
5.4
作者：
Seybert, A;Posthuma, CC;Ziebuhr, J
通讯作者：
Ziebuhr, J