Catalytically-active complex of HIV-1 integrase with a viral DNA substrate binds anti-integrase drugs

被引:30
|
作者
Alian, Akram [1 ]
Griner, Sarah L. [1 ]
Chiang, Vicki [2 ]
Tsiang, Manuel [4 ]
Jones, Gregg [4 ]
Birkus, Gabriel [4 ]
Geleziunas, Romas [4 ]
Leavitt, Andrew D. [2 ]
Stroud, Robert M. [1 ,3 ]
机构
[1] Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USA
[2] Univ Calif San Francisco, Dept Lab Med, San Francisco, CA 94158 USA
[3] Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94158 USA
[4] Gilead Sci Inc, Foster City, CA 94404 USA
基金
美国国家卫生研究院;
关键词
covalent; cross-linking; disulfide; DNA binding; strand transfer; RESOLVED FLUORESCENCE ANISOTROPY; VIRUS TYPE-1 INTEGRASE; FULL-SITE INTEGRATION; LONG TERMINAL REPEAT; IN-VITRO; RETROVIRUS INTEGRASE; CRYSTAL-STRUCTURE; DOMAIN; PROTEIN; IDENTIFICATION;
D O I
10.1073/pnas.0811919106
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
HIV-1 integration into the host cell genome is a multistep process catalyzed by the virally-encoded integrase (IN) protein. In view of the difficulty of obtaining a stable DNA-bound IN at high concentration as required for structure determination, we selected IN DNA complexes that form disulfide linkages between 5'-thiolated DNA and several single mutations to cysteine around the catalytic site of IN. Mild reducing conditions allowed for selection of the most thermodynamically-stable disulfide-linked species. The most stable complexes induce tetramer formation of IN, as happens during the physiological integration reaction, and are able to catalyze the strand transfer step of retroviral integration. One of these complexes also binds strand-transfer inhibitors of HIV antiviral drugs, making it uniquely valuable among the mutants of this set for understanding portions of the integration reaction. This novel complex may help define substrate interactions and delineate the mechanism of action of known integration inhibitors.
引用
收藏
页码:8192 / 8197
页数:6
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