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首页> 外文期刊>Journal of Virology >Regulation of Synthesis of Two Immunologically Distinct Nucleic Acid-Dependent Nucleoside Triphosphate Phosphohydrolases in Vaccinia Virus-Infected HeLa Cells
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Regulation of Synthesis of Two Immunologically Distinct Nucleic Acid-Dependent Nucleoside Triphosphate Phosphohydrolases in Vaccinia Virus-Infected HeLa Cells

机译：对痘苗病病毒感染HeLa细胞中两种免疫不同核酸依核苷三磷酸三磷酸磷酸酶的合成调节

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The two nucleic acid-dependent nucleoside triphosphate phosphohydrolases, previously purified from vaccinia virus cores, were shown to be immunologically distinct enzymes. Antiserum prepared against purified phosphohydrolase I and antiserum prepared against purified phosphohydrolase II only neutralized the activity of that enzyme used as antigen. Both enzymes were induced in HeLa cells after vaccinia infection. DNA-cellulose chromatography was used to purify the two phosphohydrolases from the cytoplasms of infected cells. The enzymes were identified by their different substrate specificities, nucleic acid dependence, and neutralization with specific antiserum. A third chromatographically separable nucleic acid-dependent phosphohydrolase similar to phosphohydrolase I in substrate specificity but not neutralizable by antiserum to either phosphohydrolase I or II, was also isolated from infected cells. No nucleic acid-dependent nucleoside triphosphate phosphohydrolase activity was detected by similar methods from uninfected HeLa cells. Formation of these virus-induced enzymes was prevented by actinomycin D and cycloheximide, indicating a requirement for de novo RNA and protein synthesis, respectively. The kinetics of induction and inhibition by cytosine arabinoside, an inhibitor of DNA synthesis, suggested that synthesis of the phosphohydrolases is a late viral function. Rifampin, an inhibitor of vaccinia virus growth which prevents virion assembly, had no inhibitory effect on the induction of the phosphohydrolases. This result was consistent with the finding that these enzymes exist in a soluble as well as in a particulate form in the cytoplasm of infected cells. Addition of another specific anti-poxviral drug, isatin-β-thiosemicarbazone, to vaccinia-infected cells partially inhibited induction of the phosphohydrolases.
机译：先前从痘苗病毒核心纯化的两种核酸依赖性核苷酸磷酸三磷酸三磷酸磷酸酶被显示为免疫不同的酶。对纯化磷酸氢酶I的抗血清制备和抗纯磷酸氢氢酶II制备的抗血清中和用作抗原的该酶的活性。在疫苗感染后，在Hela细胞中诱导了这两种酶。使用DNA-纤维素色谱法从受感染细胞的细胞质纯化两种磷酸盐酶。通过其不同的底物特异性，核酸依赖性和用特异性抗血清中和鉴定酶。从受感染的细胞中也分离出类似于底物特异性的第三种色谱分离的核酸依赖性磷酸酶，其与底物特异性，但不能通过抗血清中和磷酸盐酶I或II中和。没有通过来自未感染的HeLa细胞的类似方法检测核酸依赖性核苷三磷酸三磷酸磷酸酶活性。通过放线霉素D和环己酰亚胺预防这些病毒诱导的酶的形成，表明DE Novo RNA和蛋白质合成的要求。通过胞嘧啶阿拉伯苷的诱导和抑制的动力学，DNA合成抑制剂，表明磷酸化合物的合成是晚期病毒功能。利福平是防止病毒素组件的疫苗病毒生长的抑制剂对磷酸化合物的诱导没有抑制作用。该结果与发现这些酶存在于可溶性细胞的细胞质中的溶于溶解中的酶中的结果一致。添加另一种特异的抗痘药物，Isatin-β-硫代吡咯，对疫苗感染的细胞部分抑制磷酸盐酶的诱导。

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