Evidence that creation of invasion sites determines the rate of strand transfer mediated by HIV-1 reverse transcriptase.
Evidence that creation of invasion sites determines the rate of strand transfer mediated by HIV-1 reverse transcriptase.
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有证据表明，入侵位点的产生决定了 HIV-1 逆转录酶介导的链转移速率。
DOI：
10.1016/j.jmb.2006.08.068
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发表时间：
2006
影响因子：
5.6
通讯作者：
Bambara,RobertA
中科院分区：
文献类型：
--
作者：
Hanson,MarkNils;Balakrishnan,Mini;Roques,BernardP;Bambara,RobertA
逆转录过程中的链转移可以在人类免疫缺陷病毒1型（HIV-1）中产生基因重组，当两个不相同的基因组RNA共同包装在病毒中时。在涉及从供体到受体RNA模板的引物转换的反应中，在体外测量链转移。转移产物的动力学比供体模板上的全长合成慢得多。本研究的目的是通过定义限制其速率的步骤来更多地了解转移机制。我们先前提出的转移包括受体侵入，杂交繁殖，末端转移和受体模板上合成的重新启动的步骤。出乎意料的是，随着我们的模板的增加，受体浓度增加了转移效率，但对转移速率没有影响。具有限制杂交繁殖的同源性的短区域的模板表现出转移产物的缓慢积累，这表明对于测试的长同源性模板，杂交繁殖不是速率限制的。取代DNA受体并加入Klenow聚合酶加速了仅在DNA受体上的再起始和延伸。由于受体上更快的延伸，这导致了小的速率增加，表明在测试的RNA受体上重新引发合成不是速率限制。设计了一种底物，其中引物的5′端是单链的，并且与受体互补，即具有预制的侵入位点。对于这种底物，受体浓度的增加增加了转移速率。总之，这些数据表明，RNA酶H切割和RNA片段的解离产生侵入位点是大多数测试模板的速率限制。当存在可接近的侵入位点时，该位点的受体相互作用影响该速率。
Strand transfer during reverse transcription can produce genetic recombination in human immunodeficiency virus type 1 (HIV-1) when two genomic RNAs, that are not identical, are co-packaged in the virus. Strand transfer was measured in vitro, in reactions involving primer switching from a donor to acceptor RNA template. The transfer product appeared with much slower kinetics than full-length synthesis on the donor template. The goal of this study was to learn more about the transfer mechanism by defining the steps that limit its rate. We previously proposed transfer to include the steps of acceptor invasion, hybrid propagation, terminus transfer, and re-initiation of synthesis on the acceptor template. Unexpectedly, with our templates increasing acceptor concentration increased the transfer efficiency but had no effect on the rate of transfer. Templates with a short region of homology limiting hybrid propagation exhibited a slow accumulation of transfer products, suggesting that for tested long homology templates hybrid propagation was not rate limiting. Substituting a DNA acceptor and adding Klenow polymerase accelerated re-initiation and extension exclusively on the DNA acceptor. This lead to a small rate increase due to faster extension on the acceptor, suggesting re-initiation of synthesis on the tested RNA acceptors was not rate limiting. A substrate was designed in which the 5′ end of the primer was single stranded, and complimentary to the acceptor, i.e. having a pre-made invasion site. With this substrate, increasing concentrations of acceptor increased the rate of transfer. Together these data suggest that RNase H cleavage, and dissociation of RNA fragments creating an invasion site was rate limiting on most tested templates. When an accessible invasion site was present, acceptor interaction at that site influence the rate.
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影响因子：
2.9
作者：
J. A. Peliska;Shankar Balasubramanian;D. Giedroc;Stephen J. Benkovic
通讯作者：
J. A. Peliska;Shankar Balasubramanian;D. Giedroc;Stephen J. Benkovic
莫洛尼鼠白血病病毒逆转录过程中强终止受体模板选择所需的顺式作用元件。
影响因子：
5.6
作者：
Robert S. Topping;M. Demoitié;Nam;A. Telesnitsky
通讯作者：
A. Telesnitsky
逆转录过程中重组机制依赖于受体 RNA* 210 结构的证据
影响因子：
4.8
作者：
A. Moumen;Lucette Polomack;T. Unge;M. Véron;H. Buc;M. Negroni
通讯作者：
M. Negroni
DOI：
10.1073/pnas.89.15.6919
发表时间：
1992-08-01
影响因子：
11.1
作者：
BOYER, JC;BEBENEK, K;KUNKEL, TA
通讯作者：
KUNKEL, TA
勘误：体内抗逆转录病毒药物的较高选择压力导致 HIV-1 pol 的进化距离增加（Virology (1999) 259:1 (154-165)）
DOI：
--
发表时间：
1999
期刊：
影响因子：
--
作者：
H. Gunthard;A. Leigh;R. D’Aquila;V. Johnson;D. Kuritzkes;D. Richman;J. Wong
通讯作者：
J. Wong