CRYSTAL-STRUCTURE OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REVERSE-TRANSCRIPTASE COMPLEXED WITH DOUBLE-STRANDED DNA AT 3.0 ANGSTROM RESOLUTION SHOWS BENT DNA
CRYSTAL-STRUCTURE OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REVERSE-TRANSCRIPTASE COMPLEXED WITH DOUBLE-STRANDED DNA AT 3.0 ANGSTROM RESOLUTION SHOWS BENT DNA
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DOI：
10.1073/pnas.90.13.6320
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发表时间：
1993-07-01
影响因子：
11.1
通讯作者：
ARNOLD, E
中科院分区：
文献类型：
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作者：
JACOBOMOLINA, A;DING, JP;ARNOLD, E
人类免疫缺陷病毒1型逆转录酶（HIV-1 RT）异二聚体（P66/p51）的三元络合物的晶体结构，一种19个基准/18个基准/18个基本的双链DNA模板 - 蛋白已在3.0 Angstrom分辨率下确定。构成p66和p51聚合酶结构域的RT的四个单个子域被命名为手指，棕榈，拇指和连接[Kohlstaedt，L。A.，Wang，J.，J.，Friedman，J.M.科学256，1783-1790]。在p66和p51中，子域的总体折叠相似，但是子域的空间排列在很大程度上不同。模板播种机的A形式和B形区域被显着弯曲（40-45度）分开。最多的核酸相互作用与蛋白质相互作用主要发生在DNA的糖磷酸骨架上，并涉及p66的棕榈，拇指和手指的氨基酸残基。高度保守的区域位于聚合酶活性位点附近的p66棕榈中。这些结构元素以及p66拇指的两个α-螺旋起作用，充当夹具，可将模板启动器相对于聚合酶活性位点定位。引物末端的3'-羟基接近活性位点上催化的必不可少的ASP-1110，ASP-185和ASP-186残基，并且在接近磷酸核苷核苷的α-磷酸盐的二磷酸核苷磷酸盐的位置。 HIV-1 RT/DNA/FAB复合物的结构应有助于我们理解核酸聚合的一般机制。通过对这些研究产生的对药物抑制和抗药性的更全面了解，艾滋病疗法可能会得到增强。
The crystal structure of a ternary complex of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) heterodimer (p66/p51), a 19-base/18-base double-stranded DNA template-primer, and a monoclonal antibody Fab fragment has been determined at 3.0 angstrom resolution. The four individual subdomains of RT that make up the polymerase domains of p66 and p51 are named fingers, palm, thumb, and connection [Kohlstaedt, L. A., Wang, J., Friedman, J. M., Rice, P. A. & Steitz, T. A. (1992) Science 256, 1783-1790]. The overall folding of the subdomains is similar in p66 and p51 but the spatial arrangements of the subdomains are dramatically different. The template-primer has A-form and B-form regions separated by a significant bend (40-45-degrees). The most numerous nucleic acid interactions with protein occur primarily along the sugar-phosphate backbone of the DNA and involve amino acid residues of the palm, thumb, and fingers of p66. Highly conserved regions are located in the p66 palm near the polymerase active site. These structural elements, together with two alpha-helices of the thumb of p66, act as a clamp to position the template-primer relative to the polymerase active site. The 3'-hydroxyl of the primer terminus is close to the catalytically essential Asp-110, Asp-185, and Asp-186 residues at the active site and is in a position for nucleophilic attack on the alpha-phosphate of an incoming nucleoside triphosphate. The structure of the HIV-1 RT/DNA/Fab complex should aid our understanding of general mechanisms of nucleic acid polymerization. AIDS therapies may be enhanced by a fuller understanding of drug inhibition and resistance emerging from these studies.