Inhibition of vesicular stomatitis virus infection by spike glycoprotein. Evidence for an intracellular, G protein-requiring step.
Inhibition of vesicular stomatitis virus infection by spike glycoprotein. Evidence for an intracellular, G protein-requiring step.
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DOI：
10.1083/jcb.84.2.430
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发表时间：
1980-02
影响因子：
7.8
通讯作者：
Lenard, J
中科院分区：
文献类型：
--
作者：
Miller, D K;Lenard, J
在测定病毒指导的RNA合成的试验中，标准试验剂量的水泡性口炎病毒（VSV）对BHK细胞的感染被紫外线照射的wt VSV和TS 045（VSV的许多热不稳定的、温度敏感的G蛋白突变体之一）抑制。在45 ℃下热处理1小时后，不耐热突变体不再能够抑制VSV感染。与此相反，不耐热M蛋白突变体ts G31和非不耐热G蛋白突变体ts 044仍然可以抑制测试VSV剂量。因此，G蛋白以其天然构象的存在对于抑制感染是必要的。热处理前后ts 045或wt VSV与细胞的结合或内化至胰蛋白酶抗性状态的差异很小，并且没有证据表明细胞表面存在特异性饱和受体。在最大抑制感染的浓度下，没有一种辐照过的病毒粒子可以阻止感染性VSV与BHK细胞的结合或内化。因此，G蛋白特异性抑制作用并不发生在细胞表面，而一定发生在细胞内的某个部位，这被认为是溶酶体。溶酶体抑制剂氯喹，当加入感染病毒，完全抑制VSV感染在所有的多重感染测试，它给50%的抑制时，加入到1.5小时后感染。溶酶体在细胞内感染途径中可能的重要性进行了讨论。
In an assay measuring virus-directed RNA synthesis, infection of BHK cells by a standard test dose of vesicular stomatitis virus (VSV) was inhibited by ultraviolet light-irradiated wt VSV and by ts 045, one of a number of thermolabile, temperature-sensitive G protein mutants of VSV. After heat treatment for 1 h at 45 degrees C, the thermolabile mutants were no longer able to inhibit the VSV infection. In contrast, the thermolabile M protein mutant ts G31 and the nonthermolabile G protein mutant ts 044 could still inhibit the test VSV dose. Thus, the presence of G protein in its native conformation was necessary for inhibition of infection. There was little difference in the binding to cells or the internalization to a trypsin-resistant state of ts 045 or wt VSV before and after heat treatment, and there was no evidence of specific saturable receptors on the cell surface. None of the irradiated virions at concentrations that gave maximal inhibition of infection could prevent binding of infectious VSV to, or internalization by, BHK cells. The G protein-specific inhibition, therefore, did not occur at the cell surface but must have occurred at some intracellular site, which has been suggested to be the lysome. The lysosomal inhibitor chloroquine, when added with the infecting virus, completely inhibited VSV infection at all multiplicities of infection tested, and it gave 50% inhibition when added to 1.5 h after infection. The possible importance of the lysosome in the intracellular pathway of infection is discussed.