An unbiased proteomics approach to identify human cytomegalovirus RNA-associated proteins.
An unbiased proteomics approach to identify human cytomegalovirus RNA-associated proteins.
复制标题
一种鉴定人巨细胞病毒 RNA 相关蛋白的无偏见蛋白质组学方法。
DOI：
10.1016/j.virol.2015.02.008
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Moorman,NathanielJ
中科院分区：
文献类型：
--
作者：
Lenarcic,ErikM;Ziehr,BenjaminJ;Moorman,NathanielJ
转录后事件调节疱疹病毒基因表达，但很少疱疹病毒RNA结合蛋白已被确定。我们使用了一种无偏的方法耦合寡核苷酸（dT）亲和捕获与蛋白质组学，以确定病毒RNA相关的蛋白在感染过程中。使用这种方法，我们鉴定并确认了感染人巨细胞病毒（HCMV）的细胞中两种宿主RNA相关蛋白DHX 9和DDX 3丰度或活性的变化。我们还鉴定并证实了以前未报道的HCMV US22和pp71蛋白作为RNA相关病毒蛋白的活性，并证实了已知的病毒RNA结合蛋白pTRS 1与感染细胞中的RNA相关。此外，我们发现HCMV pp71与多聚核糖体共沉淀，与宿主和病毒RNA相关，并刺激蛋白质合成的总体速率。这些结果表明，寡聚（dT）亲和捕获与蛋白质组学相结合，提供了一种快速和直接的手段来识别RNA相关的病毒蛋白在感染过程中，可能参与基因表达的转录后控制。
Post-transcriptional events regulate herpesvirus gene expression, yet few herpesvirus RNA-binding proteins have been identified. We used an unbiased approach coupling oligo(dT) affinity capture with proteomics to identify viral RNA-associated proteins during infection. Using this approach, we identified and confirmed changes in the abundance or activity of two host RNA-associated proteins, DHX9 and DDX3, in cells infected with human cytomegalovirus (HCMV). We also identified and confirmed previously unreported activities for the HCMV US22 and pp71 proteins as RNA-associated viral proteins and confirmed that a known viral RNA-binding protein, pTRS1, associates with RNA in infected cells. Further, we found that HCMV pp71 co-sedimented with polysomes, associated with host and viral RNAs, and stimulated the overall rate of protein synthesis. These results demonstrate that oligo(dT) affinity capture coupled with proteomics provides a rapid and straightforward means to identify RNA-associated viral proteins during infection that may participate in the post-transcriptional control of gene expression.