Coronavirus Nucleocapsid Protein Facilitates Template Switching and Is Required for Efficient Transcription
Coronavirus Nucleocapsid Protein Facilitates Template Switching and Is Required for Efficient Transcription
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DOI：
10.1128/jvi.02011-09
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发表时间：
2010-02-15
影响因子：
5.4
通讯作者：
Enjuanes, Luis
中科院分区：
文献类型：
--
作者：
Zuniga, Sonia;Cruz, Jazmina L. G.;Enjuanes, Luis
如前所述，从传染性胃肠炎病毒（TGEV）纯化的核衣壳蛋白（N蛋白）增强了锤头核酶的自裂和有利的核酸退火，这些特性定义了RNA伴侣。在大肠杆菌中表达并纯化了几个TGEV n蛋白缺失突变体，并对其RNA结合能力和RNA伴侣活性进行了评价。用RNA伴侣活性分析的最小n蛋白结构域，促进DNA和RNA退火，包含中心非结构化区域（氨基酸117至268）。有趣的是，具有RNA伴侣活性的N蛋白及其缺失突变体在逆转录病毒衍生的异种系统中增强了模板切换，这加强了TGEV N蛋白是可能参与模板切换的RNA伴侣的概念。这一结果与在体内观察到的TGEV复制不需要N蛋白，但它是高效转录所必需的一致。
Purified nucleocapsid protein (N protein) from transmissible gastroenteritis virus (TGEV) enhanced hammerhead ribozyme self-cleavage and favored nucleic acid annealing, properties that define RNA chaperones, as previously reported. Several TGEV N-protein deletion mutants were expressed in Escherichia coli and purified, and their RNA binding ability and RNA chaperone activity were evaluated. The smallest N-protein domain analyzed with RNA chaperone activity, facilitating DNA and RNA annealing, contained the central unstructured region (amino acids 117 to 268). Interestingly, N protein and its deletion mutants with RNA chaperone activity enhanced template switching in a retrovirus-derived heterologous system, reinforcing the concept that TGEV N protein is an RNA chaperone that could be involved in template switching. This result is in agreement with the observation that in vivo, N protein is not necessary for TGEV replication, but it is required for efficient transcription.