Adam S A, Dreyfuss G
J Virol. 1987 Oct;61(10):3276-83. doi: 10.1128/JVI.61.10.3276-3283.1987.
The proteins that interact with cytoplasmic and nuclear polyadenylated RNA in adenovirus type 5 (Ad5) infection of HeLa cells were examined by UV-induced RNA-protein cross-linking in intact cells. The Ad5 100-kilodalton late nonvirion protein (100K protein) was cross-linked to both host and viral polyadenylated cytoplasmic RNA (mRNA). The cross-linking of the 100K protein to mRNA appears to correlate with productive infection, because the protein is not cross-linked to mRNA in abortive infection of wild-type Ad5 in monkey cells (CV-1) even though normal amounts of it are produced. However, when CV-1 cells are infected with Ad5 hr404, and Ad5 mutant which overcomes the host restriction to wild-type Ad5 infection in these cells, the 100K protein is cross-linked to mRNA. To identify and obtain antibodies to RNA-contacting proteins, a mouse was immunized with oligo(dT)-selected cross-linked RNA-protein complexes from Ad5-infected cells and the serum was used for immunoblotting experiments. It was found that in addition to the 100K protein, the Ad5 72K DNA-binding protein is also associated with RNA in the infected cells. The 72K DNA-binding protein is cross-linked to polyadenylated nuclear RNA sequences. These findings indicate that adenovirus proteins interact with RNAs in the infected cell and suggest possible mechanisms for the effects of the virus on mRNA metabolism.
通过对完整细胞进行紫外线诱导的RNA-蛋白质交联，研究了在人宫颈癌HeLa细胞中，与5型腺病毒（Ad5）感染的细胞质和细胞核多聚腺苷酸化RNA相互作用的蛋白质。Ad5 100千道尔顿晚期非病毒粒子蛋白（100K蛋白）与宿主和病毒的多聚腺苷酸化细胞质RNA（mRNA）发生交联。100K蛋白与mRNA的交联似乎与有效感染相关，因为在猴细胞（CV-1）中野生型Ad5的流产感染中，该蛋白不与mRNA交联，尽管其产生量正常。然而，当CV-1细胞感染Ad5 hr404（一种克服了这些细胞中宿主对野生型Ad5感染限制的Ad5突变体）时，100K蛋白与mRNA发生交联。为了鉴定并获得针对RNA接触蛋白的抗体，用从Ad5感染细胞中通过寡聚（dT）选择的交联RNA-蛋白质复合物免疫小鼠，并将血清用于免疫印迹实验。结果发现，除了100K蛋白外，Ad5 72K DNA结合蛋白在感染细胞中也与RNA相关联。72K DNA结合蛋白与多聚腺苷酸化的核RNA序列发生交联。这些发现表明腺病毒蛋白在感染细胞中与RNA相互作用，并提示了病毒影响mRNA代谢的可能机制。
