Peliska J A, Balasubramanian S, Giedroc D P, Benkovic S J
Department of Chemistry, Pennsylvania State University, University Park 16802.
Biochemistry. 1994 Nov 22;33(46):13817-23. doi: 10.1021/bi00250a036.
The effect of recombinant nucleocapsid protein (NCp7) from human immunodeficiency virus type 1 (HIV-1) on HIV-1 reverse transcriptase (HIV-1 RT) catalyzed DNA strand transfer reactions has been studied using kinetic methods with a defined template--primer model system. NCp7 is shown to modulate both the rate and the efficiency of DNA strand transfer synthesis. Evidence is presented that supports the role of NCp7 in catalyzing the annealing of a nascent DNA intermediate and RNA acceptor template during strand transfer. NCp7 was also found to enhance the ribonuclease H activity of HIV-1 RT and change the specificity of RNA hydrolysis, suggesting a direct role of NCp7 in HIV-1 RT catalyzed strand transfer. The implications of these findings for retroviral reverse transcription are addressed.
利用具有明确模板-引物模型系统的动力学方法,研究了来自1型人类免疫缺陷病毒(HIV-1)的重组核衣壳蛋白(NCp7)对HIV-1逆转录酶(HIV-1 RT)催化的DNA链转移反应的影响。结果表明,NCp7可调节DNA链转移合成的速率和效率。有证据支持NCp7在链转移过程中催化新生DNA中间体与RNA受体模板退火的作用。还发现NCp7增强了HIV-1 RT的核糖核酸酶H活性并改变了RNA水解的特异性,这表明NCp7在HIV-1 RT催化的链转移中起直接作用。讨论了这些发现对逆转录病毒逆转录的意义。