Role of residues 121 to 124 of vesicular stomatitis virus matrix protein in virus assembly and virus-host interaction
Role of residues 121 to 124 of vesicular stomatitis virus matrix protein in virus assembly and virus-host interaction
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DOI：
10.1128/jvi.80.8.3701-3711.2006
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发表时间：
2006-04-01
影响因子：
5.4
通讯作者：
Lyles, DS
中科院分区：
文献类型：
--
作者：
Connor, JH;McKenzie, MO;Lyles, DS
最近的解决方案的水泡性口炎病毒基质（M）蛋白的一个片段的晶体结构表明，氨基酸121至124，位于溶剂暴露环的蛋白质，是重要的M蛋白的自我协会和协会与膜。这些残基从疏水AVLA序列突变为极性序列DKQQ。从细菌中表达和纯化该突变体表明，它是结构稳定的，并且突变体M蛋白具有与野生型M蛋白相似的自缔合动力学。在感染同基因重组病毒的情况下，M蛋白的膜协会的分析表明，野生型和突变型M蛋白与膜在相同程度上。表达突变M蛋白的病毒确实显示出约三倍低的M蛋白对核衣壳-M复合物的结合亲和力。与M蛋白突变对病毒组装的相对较小的影响相反，突变病毒在MDBK而不是BHK细胞中表现出生长限制，诱导细胞凋亡较慢，病毒蛋白合成较低。尽管翻译较少的病毒蛋白，但突变病毒产生更多的病毒mRNA，表明突变病毒不能有效地促进病毒翻译。这些结果表明，VSV M蛋白的121 - 124区域在病毒装配中起次要作用，但通过增加病毒mRNA翻译参与病毒-宿主相互作用和VSV复制。
The recent solution of the crystal structure of a fragment of the vesicular stomatitis virus matrix (M) protein suggested that amino acids 121 to 124, located on a solvent-exposed loop of the protein, are important for M protein self-association and association with membranes. These residues were mutated from the hydrophobic AVLA sequence to the polar sequence DKQQ. Expression and purification of this mutant from bacteria showed that it was structurally stable and that the mutant M protein had self-association kinetics similar to those of the wild-type M protein. Analysis of the membrane association of M protein in the context of infection with isogenic recombinant viruses showed that both wild-type and mutant M proteins associated with membranes to the same extent. Virus expressing the mutant M protein did show an approximately threefold-lower binding affinity of M protein for nucleocapsid-M complexes. In contrast to the relatively minor effects of the M protein mutation on virus assembly, the mutant virus exhibited growth restriction in MDBK but not BHK cells, a slower induction of apoptosis, and lower viral-protein synthesis. Despite translating less viral protein, the mutant virus produced more viral mRNA, showing that the mutant virus could not effectively promote viral translation. These results demonstrate that the 121-to-124 region of the VSV M protein plays a minor role in virus assembly but is involved in virus-host interactions and VSV replication by augmenting viral-mRNA translation.