Site-directed mutagenesis of human immunodeficiency virus type 1 reverse transcriptase at amino acid position 138.
Site-directed mutagenesis of human immunodeficiency virus type 1 reverse transcriptase at amino acid position 138.
复制标题
人类免疫缺陷病毒1型逆转录酶第138位氨基酸的定点诱变。
作者：
H. Pelemans;A. Aertsen;K. Laethem;A. Vandamme;E. Clercq;M. Pérez;A. San;S. Velázquez;M. Camarasa;J. Balzarini
TSAO derivatives represent a class of nonnucleoside reverse transcriptase inhibitors (NNRTIs) that consistently select for the Glu138Lys resistance mutation in HIV-1 reverse transcriptase (RT). Seven RT mutants (i.e., Ala, Asp, Gln, Gly, Lys, Phe, and Tyr) were constructed by site-directed mutagenesis. The mutant Glu138Asp, Glu138Lys, Glu138Gln, Glu138Ala, and Glu138Gly RTs retained marked catalytic activity. In contrast, the Glu138Phe and Glu138Tyr RT mutants showed poor RNA-dependent DNA polymerase activity (30 and 4% of wild-type, respectively). TSAO derivatives lost their inhibitory activity against all mutant enzymes, except against the closely related Glu138Asp RT mutant that remained as sensitive to TSAOs as did wild-type RT. Other NNRTIs, including delavirdine, emivirine, and UC-781, and the NRTI ddGTP retained pronounced inhibitory activity against all mutant enzymes. When the amino acid mutations at position 138 of RT were introduced in recombinant virus clones, the sensitivity/resistance spectrum obtained toward the TSAOs and other NNRTIs was similar to those observed for the isolated recombinant mutant enzymes. The Glu138Lys RT mutant virus had the most marked resistance to TSAOs, followed by the Glu138Gln, Glu138Phe, Glu138Gly, Glu138Tyr, and Glu138Ala virus mutants. The Glu138Asp RT mutant virus kept full sensitivity to the TSAO derivatives. Mixtures of Glu138Lys RT mutant virus with the other virus clones mutated at the 138 position resulted in all cases, except for the Glu138Asp and Glu138Gly RT mutant viruses, in an outgrowth of the Glu138Lys RT mutant virus. Since the Glu138Lys RT proved most resistant to TSAO derivatives, was among the most catalytically efficient enzymes, and resulted in highly replication-competent virus, our data explain why the Glu138Lys RT mutant virus strains but not virus strains containing other amino acids at position 138 invariably emerge in cell cultures under TSAO drug pressure.
人类免疫缺陷病毒 1 型逆转录酶的 p51 亚基对于将 p66 亚基加载到模板引物上至关重要。
DOI：
10.1021/bi9728452
发表时间：
1998
期刊：
Biochemistry.
影响因子：
--
作者：
Harris,D;Lee,R;Misra,HS;Pandey,PK;Pandey,VN
通讯作者：
Pandey,VN
HIV-1 逆转录酶/核糖核酸酶 H：使用聚合酶链式反应构建的质粒在大肠杆菌中高水平表达。
DOI：
--
发表时间：
1989
期刊：
Journal of acquired immune deficiency syndromes
影响因子：
--
作者：
D'Aquila,RT;Summers,WC
通讯作者：
Summers,WC