DETERMINATION OF VIRAL-PROTEINS PRESENT IN THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PREINTEGRATION COMPLEX
DETERMINATION OF VIRAL-PROTEINS PRESENT IN THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PREINTEGRATION COMPLEX
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DOI：
10.1128/jvi.65.4.1910-1915.1991
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发表时间：
1991-04-01
影响因子：
5.4
通讯作者：
HASELTINE, WA
中科院分区：
文献类型：
--
作者：
FARNET, CM;HASELTINE, WA
用代谢放射性标记的人类免疫缺陷病毒1型病毒粒子感染细胞制备的细胞质提取物含有与标记病毒蛋白相关的病毒DNA。 通过凝胶过滤色谱和蔗糖梯度沉降，可以从这些提取物中纯化病毒DNA，作为含有整合酶的核蛋白复合物的一部分，整合酶是通过免疫沉淀和凝胶电泳分析可检测到的唯一病毒蛋白。 纯化的复合物不含可检测的gag基因产物，包括p17、p24、p7或p6，并且不含另外的pol基因产物，包括p10蛋白酶、p66和p51聚合酶或p15 RNA酶H。 几乎所有纯化的核蛋白复合物都能够在体外整合到异源DNA靶标中。 这些观察结果表明，整合酶是人类免疫缺陷病毒1型整合前复合物的一个组成部分，并建议整合酶可能是唯一的逆转录病毒DNA的整合所必需的病毒蛋白。
Cytoplasmic extracts prepared from cells infected with metabolically radiolabeled virions of human immunodeficiency virus type 1 contain viral DNA in association with labeled viral proteins. Viral DNA can be purified from these extracts by gel filtration chromatography and sucrose gradient sedimentation as a part of a nucleoprotein complex containing integrase as the only viral protein detectable by immunoprecipitation and gel electrophoretic analysis. The purified complex contains no detectable gag gene products, including p17, p24, p7, or p6, and contains no additional pol gene products, including the p10 protease, p66 and p51 polymerase, or the p15 RNase H. Nearly all of the purified nucleoprotein complexes are capable of integrating into heterologous DNA targets in vitro. These observations demonstrate that integrase is a component of the human immunodeficiency virus type 1 preintegration complex and suggest that integrase may be the only viral protein necessary for the integration of retroviral DNA.