Bukrinsky M I, Sharova N, McDonald T L, Pushkarskaya T, Tarpley W G, Stevenson M
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198-5120.
Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6125-9. doi: 10.1073/pnas.90.13.6125.
We have examined components of the preintegration complex of human immunodeficiency virus type 1 (HIV-1) and have analyzed features which govern the association of these components. HIV-1 nucleoprotein complexes, isolated from nuclear and cytoplasmic extracts of CD4+ cells after acute virus infection, contained viral RNA and DNA in association with viral matrix (MA), integrase (IN), and reverse transcriptase (RT) antigens but not capsid (CA) antigens and possessed integration activity in vitro. Association of IN but not RT or MA antigens with viral DNA was detergent-stable. Analysis of viral DNA synthesis and nuclear import of viral nucleoprotein complexes in the presence of a reversible RT inhibitor demonstrated that reverse transcription of viral RNA could be completed entirely in the host cell nucleus. Our studies demonstrate structural and functional features of the nucleoprotein (preintegration) complex of HIV-1 which are pertinent to the understanding of early events in the lentiviral life cycle.
我们检测了1型人类免疫缺陷病毒(HIV-1)整合前复合物的组成成分,并分析了决定这些成分相互结合的特征。急性病毒感染后,从CD4+细胞的核提取物和细胞质提取物中分离出的HIV-1核蛋白复合物,含有与病毒基质(MA)、整合酶(IN)和逆转录酶(RT)抗原相结合的病毒RNA和DNA,但不含有衣壳(CA)抗原,并且在体外具有整合活性。IN抗原而非RT或MA抗原与病毒DNA的结合对去污剂稳定。在存在可逆性RT抑制剂的情况下,对病毒DNA合成和病毒核蛋白复合物的核输入进行分析表明,病毒RNA的逆转录可在宿主细胞核中完全完成。我们的研究揭示了HIV-1核蛋白(整合前)复合物的结构和功能特征,这些特征与理解慢病毒生命周期中的早期事件相关。