Interaction of the reverse transcriptase of human immunodeficiency virus type 1 with DNA.
Interaction of the reverse transcriptase of human immunodeficiency virus type 1 with DNA.
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1 型人类免疫缺陷病毒逆转录酶与 DNA 的相互作用。
作者：
M. Bakhanashvili;A. Hizi
在DNA合成期间，人类免疫缺陷病毒（HIV）逆转录酶（RT）与模板引物的结合先于其与核苷酸三磷酸的结合。用凝胶阻滞法研究了寡核苷酸DNA与HIV-1 RT的相互作用。HIV-1 RT的同源二聚体（p66/p66）和异源二聚体（p66/p51）亚型都能够结合DNA寡聚体。因此，HIV-1 RT与DNA相互作用的所有进一步研究都是用异二聚体RT进行的。我们研究了最佳结合的条件。RT-DNA复合物的形成不依赖于引物，单链和双链DNA（平端或凹陷）的DNA结合程度是不可区分的。发现RT的DNA结合活性依赖于寡核苷酸长度。HIV-1 RT结合DNA，没有明显的序列特异性。因此，这种酶属于序列非特异性DNA结合蛋白。发现这种相互作用不依赖于DNA合成。RT-DNA复合物的形成不受模板互补或非互补dNTP的存在的影响，这表明DNA聚合和RT与dNTP的结合都不影响复合物的稳定性。凝胶阻滞测定也用于检查各种HIV-1 RT抑制剂（即，AZT-TP、ddTTP、TIBO和3，5，8-三羟基-4-喹诺酮）对酶-DNA相互作用的影响。结果表明，这些化合物的作用模式的差异。（250字处删节）
During DNA synthesis, the binding of human immunodeficiency virus (HIV) reverse transcriptase (RT) to the template-primer precedes its binding to nucleotide triphosphates. The interaction of oligonucleotide DNA with HIV-1 RT was investigated by using a gel retardation assay. Both homodimeric (p66/p66) and heterodimeric (p66/p51) isoforms of HIV-1 RT were capable of binding the DNA oligomers. Thus, all further studies on the interaction of HIV-1 RT with DNA were done with heterodimeric RT. We have studied the conditions for optimal binding. The formation of the RT-DNA complex was primer-independent, and the extent of DNA binding was indistinguishable for both single-stranded and double-stranded DNA (either blunt-ended or recessed). The DNA binding activity of the RT was found to be dependent on oligonucleotide length. HIV-1 RT binds DNA with no apparent sequence specificity. Hence, this enzyme belongs to the sequence nonspecific DNA binding proteins. The interaction was found to be independent of DNA synthesis. The formation of the RT-DNA complex was not influenced by the presence of either template-complementary or noncomplementary dNTPs, indicating that neither DNA polymerization nor binding of the RT to the dNTP affects the stability of the complex. The gel retardation assay was utilized to examine also the effect of various HIV-1 RT inhibitors (i.e., AZT-TP, ddTTP, TIBO, and 3,5,8-trihydroxy-4-quinolone) on the enzyme-DNA interaction. The results indicate differences in the modes of action of these compounds.(ABSTRACT TRUNCATED AT 250 WORDS)
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DNA 聚合酶 δ、增殖细胞核抗原和合成寡核苷酸模板引物的相互作用。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Ng,L;McConnell,M;Tan,CK;Downey,KM;Fisher,PA
通讯作者：
Fisher,PA
引物结合位点的特性以及镁离子在 KB 细胞 DNA 聚合酶 α 的引物模板识别中的作用。
影响因子：
2.9
作者：
Fisher,PA;Korn,D
通讯作者：
Korn,D
DOI：
10.1073/pnas.90.13.6320
发表时间：
1993-07-01
影响因子：
11.1
作者：
JACOBOMOLINA, A;DING, JP;ARNOLD, E
通讯作者：
ARNOLD, E
DOI：
10.1073/pnas.86.21.8343
发表时间：
1989-11-01
影响因子：
11.1
作者：
PERRINO, FW;PRESTON, BD;LOEB, LA
通讯作者：
LOEB, LA
影响因子：
56.9
作者：
KOHLSTAEDT, LA;WANG, J;STEITZ, TA
通讯作者：
STEITZ, TA