Correlation of RNA binding affinity of avian oncornavirus p19 proteins with the extent of processing of virus genome RNA in cells.
Correlation of RNA binding affinity of avian oncornavirus p19 proteins with the extent of processing of virus genome RNA in cells.
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禽冠状病毒 p19 蛋白的 RNA 结合亲和力与细胞中病毒基因组 RNA 加工程度的相关性。
DOI：
10.1128/jvi.35.3.722-731.1980
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发表时间：
1980
影响因子：
5.4
通讯作者：
Smith,RE
中科院分区：
文献类型：
--
作者：
Leis,JP;Scheible,P;Smith,RE
我们从Rous肉瘤病毒、禽成髓细胞增生症病毒、B77肉瘤病毒、成髓细胞增殖症相关病毒2(0)和PR-E 95-C病毒布拉格C株中纯化了p19蛋白，并用硝酸纤维素膜结合技术测定了它们与60S病毒RNA的结合亲和力。Rous肉瘤病毒布拉格C、禽成髓细胞增生症病毒和B77肉瘤病毒的p19蛋白与同源和异源60S RNA的表观结合常数相似(1.5×10(11)~2.6×10(11)L/mol)，而成髓细胞相关病毒2(0)和PR-E 95-C病毒的p19蛋白的表观结合常数则低10倍。通过在含有甲基氢氧化汞的琼脂糖凝胶上分离、转移到重氮苯氧甲基纸片上并与70个核苷酸互补的DNA探针杂交，测定了感染Rous肉瘤病毒布拉格C、成髓细胞相关病毒2(0)和PR-E 95-C病毒的细胞质中含有病毒特异性多腺苷酸的RNA的大小和相对数量。在感染Rous肉瘤病毒布拉格C的细胞中，我们检测到3.4×10(6)-、1.9×10(6)-和1.1×10(6)-Dalton RNA，在PR-E 95-C病毒感染的细胞中，我们检测到3.4×10(6)-、1.9×10(6)-和0.7×10(6)-Dalton RNA，在感染成髓细胞相关病毒-2(0)的细胞中，我们检测到3×10(6)-和1.3×10(6)-Dalton RNA。这些RNA物种中的每一个都含有来自基因组长度RNA 5‘端的RNA序列，与5’70核苷酸互补DNA的杂交证明了这一点。在感染髓母细胞增殖症相关病毒2(0)和PR-E 95-C病毒的细胞中，亚基因组mRNA与基因组长度的RNA的比率比感染Rous肉瘤病毒布拉格C的细胞高三到五倍。这些结果表明，感染细胞中病毒RNA的加工较多与p19蛋白与病毒RNA的结合亲和力较低有关，这与p19蛋白控制细胞中病毒RNA加工的假设是一致的。
We purified the p19 proteins from the Prague C strain of Rous sarcoma virus, avian myeloblastosis virus, B77 sarcoma virus, myeloblastosis-associated virus-2(0), and PR-E 95-C virus and measured their binding affinities for 60S viral RNA by the nitrocellulose filter binding technique. The apparent association constants of the p19 proteins from Rous sarcoma virus Prague C, avian myeloblastosis virus, and B77 sarcoma virus for homologous and heterologous 60S RNAs were similar (1.5 x 10(11) to 2.6 x 10(11) liters/mol), whereas those of myeloblastosis-associated virus-2(0) and PR-E 95-C virus were 10-fold lower. The sizes and relative amounts of the virus-specific polyadenylic acid-containing RNAs in the cytoplasms of cells infected with Rous sarcoma virus Prague C, myeloblastosis-associated virus-2(0), and PR-E 95-C virus were determined by fractionating the RNAs on agarose gels containing methylmercury hydroxide, transferring them to diazobenzyloxymethyl paper and hybridizing them to a 70-nucleotide complementary DNA probe. In cells infected with Rous sarcoma virus Prague C we detected 3.4 x 10(6)-, 1.9 x 10(6)-, and 1.1 x 10(6)-dalton RNAs, in PR-E 95-C virus-infected cells we detected 3.4 x 10(6)-, 1.9 x 10(6)- and 0.7 x 10(6)-dalton RNAs, and in cells infected with myeloblastosis-associated virus-2(0) we detected 3 x 10(6)- and 1.3 x 10(6)-dalton RNAs. Each of these RNA species contained RNA sequences derived from the 5' terminus of genome-length RNA, as evidenced by hybridization with the 5' 70-nucleotide complementary DNA. The ratios of subgenomic mRNA's to genome-length RNAs in cells infected with myeloblastosis-associated virus-2(0) and PR-E 95-C virus were three- to five-fold higher than the ratio in cells infected with Rous sarcoma virus Prague C. These results suggest that more processing of viral RNA in infected cells is correlated with lower binding affinities of the p19 protein for viral RNA, and they are consistent with the hypothesis that the p19 protein controls processing of viral RNA in cells.