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Expression and function of structural proteins of paramyxovirus transcription complex

副粘病毒转录复合物结构蛋白的表达和功能

基本信息

批准号：
07680736
负责人：
MATSUMURA Haruo
金额：
$ 0.64万
依托单位：
Kinki University (1996-1997)Mie University (1995)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

V gene of human parainfluenza virus type 2 (hPIV2) , a member of paramyxoviruses, is transcribed into both a V and a P mRNA.The V mRNA is a faithful transcript of the V gene, however the P mRNA is transcribed by RNA editing mechanism in hPIV2-infected mammalian cells. Thus both the P and the V proteins are translated in the cells. We constructed recombinant baculoviruses containing the original V gene, which has seven G residues at its editing site and a manipulated V gene, which has ten G residues at its editing site. Unexpectedly, a small amount of the P protein was synthesized in addition to the V protein when the V gene was expressed in the recombinant baculovirus-infected insect cells. Furthermore, synthesis of the P protein increased when a recombinant baculovirus containing a V gene, which has extended length of G residue atretch, that is ten G residues, at its editing site, was expressed in the insect cells. Both the P and the V proteins were synthesized in in vitro translation of mRNA from the recombinant baculovirus-infected cells. Moreover, both G-residue insertions and a deletion were detected in mRNA.While the P protein was not detected in in vitro translation of the V RNA that was transcribed in vitro by T7 RNA polymerase. These results suggest that the non-templated G residues were inserted and deleted at the transcription level without paramyxovirus L protein.

人副流感病毒 2 型 (hPIV2) 是副粘病毒的一员，其 V 基因被转录为 V 和 P mRNA。V mRNA 是 V 基因的忠实转录物，但 P mRNA 在 hPIV2 感染的哺乳动物细胞中通过 RNA 编辑机制转录。因此P和V蛋白都在细胞中翻译。我们构建了含有原始 V 基因（其编辑位点有 7 个 G 残基）和操纵的 V 基因（其编辑位点有 10 个 G 残基）的重组杆状病毒。出乎意料的是，当V基因在重组杆状病毒感染的昆虫细胞中表达时，除了V蛋白之外，还合成了少量的P蛋白。此外，当含有V基因的重组杆状病毒在昆虫细胞中表达时，P蛋白的合成增加，该V基因在其编辑位点具有延长的G残基长度，即10个G残基。 P 和 V 蛋白都是在重组杆状病毒感染细胞的 mRNA 体外翻译中合成的。此外，在mRNA中检测到G残基插入和缺失。而在由T7 RNA聚合酶体外转录的V RNA的体外翻译中未检测到P蛋白。这些结果表明，在没有副粘病毒L蛋白的情况下，非模板化的G残基在转录水平上被插入和删除。