Poncet D, Aponte C, Cohen J
Laboratoire de Virologie et Immunologie Moléculaires, Institut National de la Recherche Agronomique, Jouy en Josas, France.
J Virol. 1993 Jun;67(6):3159-65. doi: 10.1128/JVI.67.6.3159-3165.1993.
Interaction between viral proteins and RNAs has been studied in rotavirus-infected cells. The use of UV cross-linking followed by immunoprecipitation and labeling with T4 polynucleotide kinase allowed us to detect interactions between RNA and nonstructural viral proteins. The RNAs linked to the nonstructural protein NSP3 have been identified as rotavirus mRNAs, and the sequences of the RNase T1-protected fragments have been established. These sequences correspond to the 3' end sequence common to all rotavirus group A genes. We also show that the last 3' nucleotide is cross-linked to the protein and that monomeric and multimeric forms of NSP3 are bound to rotavirus mRNA. The role of NSP3 in rotavirus replication is discussed in the light of our results and by comparison with other RNA-binding proteins of members of the Reoviridae family.
已在轮状病毒感染的细胞中研究了病毒蛋白与RNA之间的相互作用。通过紫外线交联，随后进行免疫沉淀并用T4多核苷酸激酶标记，使我们能够检测RNA与非结构病毒蛋白之间的相互作用。与非结构蛋白NSP3相连的RNA已被鉴定为轮状病毒mRNA，并且已确定了核糖核酸酶T1保护片段的序列。这些序列对应于所有A组轮状病毒基因共有的3'末端序列。我们还表明，最后一个3'核苷酸与该蛋白交联，并且NSP3的单体和多聚体形式与轮状病毒mRNA结合。根据我们的结果并与呼肠孤病毒科成员的其他RNA结合蛋白进行比较，讨论了NSP3在轮状病毒复制中的作用。
