Protein synthesis in lysates of Aedes albopictus cells infected with vesicular stomatitis virus.
Protein synthesis in lysates of Aedes albopictus cells infected with vesicular stomatitis virus.
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感染水疱性口炎病毒的白纹伊蚊细胞裂解物中的蛋白质合成。
DOI：
10.1128/mcb.2.10.1174-1186.1982
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发表时间：
1982
影响因子：
5.3
通讯作者：
Stollar,V
中科院分区：
文献类型：
--
作者：
Gillies,S;Stollar,V
白纹伊蚊细胞（克隆LT-C7）在感染水疱性口炎病毒（VSV）后表现出明显的细胞病变作用和蛋白合成抑制（宿主和病毒），但前提是：(i)培养在34°C而不是28°C孵育，（ii）培养基（S. Gillies和V. Stollar, Mol. Cell）中存在血清。biol.2:66 - 75, 1982)。了解vsv感染的da如何关闭蛋白质合成。我们比较了vsv感染细胞和对照细胞提取物制备的无细胞蛋白合成。vsv感染细胞在34°C下保存6小时后提取的提取物在血清中至少反映了在完整细胞中观察到的两个方面：(i)它们显示出进行蛋白质合成的能力明显减弱（无论是由内源性还是外源性添加的mRNA引导），（ii）在体外，特定核糖体蛋白的[γ-32P]ATP磷酸化减少（Gillies和Stollar， Mol. Cell）。biol.2:66 - 75, 1982)。此外，与起始水平上的阻滞一致，通过VSV 12与18S mRNA的结合测量，无活性提取物中80S起始复合物的形成减少。将未感染细胞的S-100组分加入到失活提取物中，可以逆转上述每种变化；即恢复了蛋白质合成活性，刺激了80S起始复合物的形成，增加了上述特定核糖体蛋白的磷酸化。S-100馏分中的活性成分是热不稳定的，不透析性的，在硫酸铵对S-100馏分进行分馏后，在饱和度为40 ~ 70%的馏分中发现了活性成分。我们的发现提示VSV感染a。白纹珠细胞通过灭活S-100片段中的大分子组分（可能是蛋白质）来抑制蛋白质合成，该组分可能参与蛋白质合成的起始。更具体地说，我们认为该成分参与了核糖体亚基的连接，形成80S起始复合物。
Aedes albopictuscells (clone LT-C7) showed a marked cytopathic effect and inhibition of protein synthesis (both host and viral) after infection with vesicular stomatitis virus (VSV), but only if (i) cultures were incubated at 34°C rather than 28°C and (ii) serum was present in the medium (S. Gillies and V. Stollar, Mol. Cell. Biol.2:66–75, 1982). To learn more about how protein synthesis is shut off in VSV-infectedA. albopictuscells, we have compared cell-free protein synthesis in extracts prepared from VSV-infected cells and control cells. Extracts prepared 6 h after infection from VSV-infected cells maintained at 34°C in the presence of serum reflected what was observed with intact cells in at least two respects: (i) they showed a markedly diminished capacity to carry out protein synthesis (whether directed by endogenous or exogenously added mRNA), and (ii) there was decreased phosphorylation in vitro by [γ-32P]ATP of a specific ribosomal protein (Gillies and Stollar, Mol. Cell. Biol.2:66–75, 1982). In addition, and consistent with a block at the level of initiation, the formation of 80S initiation complexes, as measured by binding of VSV 12 to 18S mRNA, was reduced in the inactive extracts. Addition of an S-100 fraction from uninfected cells to the inactive extract reversed each of the aforementioned changes; i.e., it restored protein synthetic activity, it stimulated the formation of 80S initiation complexes, and it increased phosphorylation of the specific ribosomal protein referred to above. The active component in the S-100 fraction was heat labile and non-dialyzable and, upon ammonium sulfate fractionation of the S-100 fraction, was found in the 40 to 70% saturation fraction. Our findings suggest that VSV infection ofA. albopictuscells inhibits protein synthesis by inactivating a macromolecular component, probably a protein, in the S-100 fraction which may be involved in the initiation of protein synthesis. More specifically, we suggest that this component is involved in the joining of the ribosomal subunits to form 80S initiation complexes.