Characterization of intracellular reverse transcription complexes of human immunodeficiency virus type 1
Characterization of intracellular reverse transcription complexes of human immunodeficiency virus type 1
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DOI：
10.1128/jvi.75.8.3626-3635.2001
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发表时间：
2001-04-01
影响因子：
5.4
通讯作者：
Goff, SP
中科院分区：
文献类型：
--
作者：
Fassati, A;Goff, SP
为了研究人类免疫缺陷病毒1型（HIV-1）生命周期的早期事件，我们分析了从急性感染细胞中分离的介导逆转录的细胞内复合物。部分纯化的逆转录复合物（RTCs）的平衡密度分级和速度沉降表明，两个物种的RTCs形成，但只有一个物种是能够合成DNA。大多数的衣壳，基质，和逆转录酶（RT）蛋白解离后不久，细胞感染的复合物，但Vpr仍然与RTC。感染后1，4和7小时分离的RTCs能够在体外进行逆转录，这表明一小部分RT仍然与它们相关。感染后早期分离的HIV RTC的沉降速度约为560 S。之后，出现了不同的沉积物种，其沉积速率在350 - 100 S之间。核相关RTC的沉降速度为80 S。逆转录开始后不久，RTC内的病毒强终止DNA对核酸酶消化敏感，并且在逆转录几乎完成时受到保护。
To examine the early events of the life cycle of human immunodeficiency virus type 1 (HIV-1), we analyzed the intracellular complexes mediating reverse transcription isolated from acutely infected cells. Partial purification of the reverse transcription complexes (RTCs) by equilibrium density fractionation and velocity sedimentation indicated that two species of RTCs are formed but only one species is able to synthesize DNA. Most of the capsid, matrix, and reverse transcriptase (RT) proteins dissociate from the complex soon after cell infection, but Vpr remains associated with the RTC. The RTCs isolated 1, 4, and 7 h after infection are competent for reverse transcription in vitro, indicating that a small proportion of RT remains associated with them. HIV RTCs isolated early after infection have a sedimentation velocity of approximately 560S. Later, different species with a sedimentation velocity ranging from 350S to 100S appear. Nuclear-associated RTCs have a sedimentation velocity of 80S, Shortly after initiation of reverse transcription, the viral strong-stop DNA within the RTC is sensitive to nuclease digestion and becomes protected when reverse transcription is almost completed.