Mutational analysis of two conserved sequence motifs in HIV-1 reverse transcriptase.
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, aspartates 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 resulted in reduced reverse trascriptase activity but the mutation of tyrosine-183 to serine caused a significant increase in the Km for dTTP and the Ki for inhibition by 3'-azidothymidine-triphosphate, 2',3'-dideoxythymidine-triphosphate and phosphonoformic acid.
Lowe
D M
DM
Department of Molecular Sciences, Wellcome Research Laboratories, Beckenham, Kent, U.K.
Parmar
V
V
Kemp
S D
SD
Larder
B A
BA
D000595:Amino Acid Sequence; D004252:DNA Mutational Analysis; D054306:Dideoxynucleotides; D017245:Foscarnet; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D066298:In Vitro Techniques; D007700:Kinetics; D008969:Molecular Sequence Data; D010746:Phosphonoacetic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; D013329:Structure-Activity Relationship; D013942:Thymine Nucleotides / Q000494:pharmacology; D015215:Zidovudine / Q000031:analogs & derivatives / Q000494:pharmacology
D054306:Dideoxynucleotides; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D017245:Foscarnet; D015215:Zidovudine; C052873:zidovudine triphosphate; D012194:RNA-Directed DNA Polymerase; C017216:2',3'-dideoxythymidine triphosphate; D010746:Phosphonoacetic Acid