Identification and molecular characterization of the RNA polymerase-binding motif of infectious bursal disease virus inner capsid protein VP3
Identification and molecular characterization of the RNA polymerase-binding motif of infectious bursal disease virus inner capsid protein VP3
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DOI：
10.1128/jvi.77.4.2459-2468.2003
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发表时间：
2003-02-01
影响因子：
5.4
通讯作者：
Lombardo, E
中科院分区：
文献类型：
--
作者：
Maraver, A;Clemente, R;Lombardo, E
传染性法氏囊病病毒（IBDV）是双核糖核酸病毒科的一种病毒，是引起禽类重要传染病的病原。IBDV分子生物学的主要方面，如组装和复制，至今知之甚少。我们以前已经表明，假定的病毒编码的蛋白质的水解。RNA依赖性RNA聚合酶VP 1通过其与内衣壳蛋白VP 3的相互作用介导。在这里，我们报告的VP 1-VP 3相互作用的表征。RNA酶A处理的VP 1-和VP 3-含有提取物不影响VP 1-VP 3复合物的形成，表明复合物的形成需要建立蛋白质-蛋白质相互作用。使用一组VP 3缺失突变体允许在VP 3的C末端上的高度带电的16个氨基酸延伸内绘制VP 3的VP 1结合基序。当与不相关的蛋白融合时，VP 3的该区域足以赋予VP 1结合活性。此外，对应于VP 3的VP 1结合区的肽特异性抑制VP 1-VP 3复合物的形成。在IBDV感染的细胞中含有VP 1结合基序的特洛伊肽的存在特异性地减少了感染性病毒的产生，从而表明VP 1-VP 3复合物的形成在IBDV复制中起关键作用。
Infectious bursal disease virus (IBDV), a member of the Birnaviridae family, is the causative agent of one of the most important infectious poultry diseases. Major aspects of the molecular biology of IBDV, such as assembly and replication, are as yet poorly understood. We have previously shown that encapsidation of the putative virus-encoded. RNA-dependent RNA polymerase VP1 is mediated by its interaction with the inner capsid protein VP3. Here, we report the characterization of the VP1-VP3 interaction. RNase A treatment of VP1- and VP3-containing extracts does not affect the formation of VP1-VP3 complexes, indicating that formation of the complex requires the establishment of protein-protein interactions. The use of a set of VP3 deletion mutants allowed the mapping of the VP1 binding motif of VP3 within a highly charged 16-amino-acid stretch on the C terminus of VP3. This region of VP3 is sufficient to confer VP1 binding activity when fused to an unrelated protein. Furthermore, a peptide corresponding to the VP1 binding region of VP3 specifically inhibits the formation of VP1-VP3 complexes. The presence of Trojan peptides containing the VP1 binding motif in IBDV-infected cells specifically reduces infective virus production, thus showing that formation of VP1-VP3 complexes plays a critical role in IBDV replication.