Mutational analysis of two conserved sequence motifs in HIV‐1 reverse transcriptase

FEBS Letters

Kemp, Sharon D.¹  Larder, Brendan A.¹  Lowe, Denise M.¹  Parmar, Vanita¹ 

[1] Department of Molecular Sciences, The Wellcome Research Laboratories, Langley Court, Beckenham, Kent BR3 3BS, UK

关键词: Reverse transcriptase (HIV-1);    Site-directed mutagenesis;    Conserved sequence motif;    Enzyme inhibition;    RT;    reverse transcriptase;    AZT-TP;    3′-azidothymidine triphosphate;    ddTTP;    2′;    3′-dideoxythymidine triphosphate;   

DOI  :  10.1016/0014-5793(91)80484-K

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Two conserved sequence motifs, occurring in HIV-1 reverse transcriptase at residues 110–116 and 183–190, have been studied using site-directed mutagenesis of the cloned gene. In particular, aspariates at positions 185 and 186 have each been mutated to either asparagine or glutamate. The resulting mutant proteins were catalytically inactive but still able to bind the template-primer complex, poly rA-oligo dT. Other mutations in these regions results in reduced reverse transcriptase activity but the mutation of tyrosine-183 to serine caused a significant increase in the K_m for dTTP and the K_m for inhibition by 3′-azi-Jothymidine-triphosphate, 2′3′-dideoxythymidine-triphosphate and phosphonoformic acid.

【 授权许可】

Unknown