Garcin D, Lezzi M, Dobbs M, Elliott R M, Schmaljohn C, Kang C Y, Kolakofsky D
Department of Genetics and Microbiology, University of Geneva School of Medicine, Switzerland.
J Virol. 1995 Sep;69(9):5754-62. doi: 10.1128/JVI.69.9.5754-5762.1995.
We examined the 5' ends of Hantaan virus (HTN) genomes and mRNAs to gain insight into the manner in which these chains were initiated. Like those of all members of the family Bunyaviridae described so far, the HTN mRNAs contained 5' terminal extensions that were heterogeneous in both length and sequence, presumably because HTN also "cap snatches" host mRNAs to initiate the viral mRNAs. Unexpectedly, however, almost all of the mRNAs contained a G residue at position -1, and a large fraction also lacked precisely one of the three UAG repeats at the termini. The genomes, on the other hand, commenced with a U residue at position +1, but only 5' monophosphates were found here, indicating that these chains may not have initiated with UTP at this position. Taken together, these unusual findings suggest a prime-and-realign mechanism of chain initiation in which mRNAs are initiated with a G-terminated host cell primer and genomes with GTP, not at the 3' end of the genome template but internally (opposite the template C at position +3), and after extension by one or a few nucleotides, the nascent chain realigns backwards by virtue of the terminal sequence repeats, before processive elongation takes place. For genome initiation, an endonuclease, perhaps that involved in cap snatching, is postulated to remove the 5' terminal extension of the genome, leaving the 5' pU at position +1.
我们研究了汉坦病毒(HTN)基因组和mRNA的5'末端,以深入了解这些链的起始方式。与迄今为止描述的布尼亚病毒科所有成员一样,HTN mRNA含有5'末端延伸,其长度和序列均异质,推测是因为HTN也“抢夺”宿主mRNA来起始病毒mRNA。然而,出乎意料的是,几乎所有的mRNA在-1位置都含有一个G残基,并且很大一部分在末端也恰好缺少三个UAG重复序列中的一个。另一方面,基因组在+1位置以U残基开始,但在此处仅发现5'单磷酸,这表明这些链可能不是在此位置以UTP起始的。综合起来,这些不寻常的发现提示了一种引物与重排的链起始机制,即mRNA以G末端的宿主细胞引物起始,基因组以GTP起始,不是在基因组模板的3'末端,而是在内部(与+3位置的模板C相对),并且在延伸一个或几个核苷酸后,新生链凭借末端序列重复向后重排,然后进行持续延伸。对于基因组起始,推测一种内切核酸酶,可能是参与抢夺帽结构的那种,会去除基因组的5'末端延伸,在+1位置留下5' pU。