Roles of serine and threonine residues of mumps virus P protein in viral transcription and replication.
Roles of serine and threonine residues of mumps virus P protein in viral transcription and replication.
复制标题
腮腺炎病毒P蛋白丝氨酸和苏氨酸残基在病毒转录和复制中的作用。
DOI：
10.1128/jvi.03673-13
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发表时间：
2014
影响因子：
5.4
通讯作者：
He,Biao
中科院分区：
文献类型：
--
作者：
Pickar,Adrian;Xu,Pei;Elson,Andrew;Li,Zhuo;Zengel,James;He,Biao
腮腺炎病毒（MuV）是一种含有负义非节段RNA基因组的副粘病毒，是一种引起急性感染的人类病原体，症状范围从腮腺炎到轻度脑膜炎和严重脑炎。接种腮腺炎病毒疫苗对减少腮腺炎病例有效。然而，最近在接种疫苗的人群中发生了大规模疫情。没有抗MuV药物。了解MuV的复制可能会导致新的抗病毒策略。MuV RNA依赖性RNA聚合酶最少由磷蛋白（P）和大蛋白（L）组成。P蛋白被严重磷酸化。为了研究P的丝氨酸（S）和苏氨酸（T）残基在病毒RNA转录和复制中的作用，对P进行质谱和突变分析。P是一种由392个氨基酸残基组成的蛋白质，含有64个S和T残基。我们已经发现，在微型基因组系统中，突变9个S/T残基显著降低活性，并且将101处的残基T突变为A（T101 A）显著增强活性。回收并分析含有P-T101 A突变的重组病毒（rMuV-P-T101 A）。rMuV-P-T101 A生长至更高滴度并且在早期时间点具有增加的蛋白质表达。总之，这些结果表明MuV-P-T101的磷酸化在病毒RNA合成中起负面作用。这是第一次系统地分析了副粘病毒P蛋白的S/T残基，S/T残基对病毒RNA合成至关重要。重要的是，Mumps病毒（MuV）是一种重新出现的副粘病毒，在疫苗接种覆盖率非常高的美国引起了大规模爆发。没有抗MuV药物。本研究系统地分析了MuV P的Ser/Thr残基在病毒RNA合成中的作用。我们已经确定了P的S/T残基对MuV RNA合成和磷酸化位点是重要的病毒RNA合成的关键。这项工作导致更好地了解病毒RNA的合成，以及潜在的新战略，以控制腮腺炎。
Mumps virus (MuV), a paramyxovirus containing a negative-sense nonsegmented RNA genome, is a human pathogen that causes an acute infection with symptoms ranging from parotitis to mild meningitis and severe encephalitis. Vaccination against mumps virus has been effective in reducing mumps cases. However, recently large outbreaks have occurred in vaccinated populations. There is no anti-MuV drug. Understanding replication of MuV may lead to novel antiviral strategies. MuV RNA-dependent RNA polymerase minimally consists of the phosphoprotein (P) and the large protein (L). The P protein is heavily phosphorylated. To investigate the roles of serine (S) and threonine (T) residues of P in viral RNA transcription and replication, P was subjected to mass spectrometry and mutational analysis. P, a 392-amino acid residue protein, has 64 S and T residues. We have found that mutating nine S/T residues significantly reduced and mutating residue T at 101 to A (T101A) significantly enhanced activity in a minigenome system. A recombinant virus containing the P-T101A mutation (rMuV-P-T101A) was recovered and analyzed. rMuV-P-T101A grew to higher titers and had increased protein expression at early time points. Together, these results suggest that phosphorylation of MuV-P-T101 plays a negative role in viral RNA synthesis. This is the first time that the P protein of a paramyxovirus has been systematically analyzed for S/T residues that are critical for viral RNA synthesis.IMPORTANCEMumps virus (MuV) is a reemerging paramyxovirus that caused large outbreaks in the United States, where vaccination coverage is very high. There is no anti-MuV drug. In this work, we have systematically analyzed roles of Ser/Thr residues of MuV P in viral RNA synthesis. We have identified S/T residues of P critical for MuV RNA synthesis and phosphorylation sites that are important for viral RNA synthesis. This work leads to a better understanding of viral RNA synthesis as well as to potential novel strategies to control mumps.