Chimeric human immunodeficiency virus type 1 and feline immunodeficiency virus reverse transcriptases: role of the subunits in resistance/sensitivity to non-nucleoside reverse transcriptase inhibitors.
Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are specific for human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and do not inhibit HIV-2. Given that the amino acids lining the NNRTI-specific pocket of HIV-1 RT display higher similarity to the corresponding feline immunodeficiency virus (FIV) RT amino acids than to HIV-2 RT, the susceptibility of FIV RT and chimeric HIV-1/FIV RTs to NNRTIs and the role of the p51 subunit in the inhibitory action of NNRTIs were investigated. We found that the wild-type FIV RT and the FIVp66/HIVp51 chimeric enzyme showed no susceptibility for NNRTIs. On the other hand, the chimeric HIVp66/FIVp51 RT retained a sensitivity spectrum for NNRTIs similar to that of the wild-type HIV-1 RT. The noncompetitive nature of inhibition of HIV-1 RT by nevirapine was also observed with the HIVp66/FIVp51 chimeric enzyme. Inhibition of the chimeric RTs by nucleoside reverse transcriptase inhibitors and foscarnet was in the same range as observed for the corresponding HIVp66/HIVp51 and FIVp66/FIVp51 wild-type enzymes. The chimeric RTs had an affinity (K(m)) for their dNTP substrate and template/primer comparable with that of the wild-type HIV-1 and FIV RTs, but their catalytic efficacy (k(cat)) was markedly decreased. This decreased catalytic efficacy of the RT chimeras may suggest suboptimal interactions between p66 and p51 in the chimeric enzymes. Our results point to a minor role of the p51 subunit in the sensitivity to RT inhibitors.
Auwerx
Joeri
J
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Leuven, Belgium.
North
Thomas W
TW
Preston
Bradley D
BD
Klarmann
George J
GJ
De Clercq
Erik
E
Balzarini
Jan
J
D000595:Amino Acid Sequence; D003848:Deoxyguanine Nucleotides / Q000737:chemistry / Q000494:pharmacology; D054306:Dideoxynucleotides; D004352:Drug Resistance, Microbial / Q000502:physiology*; D004926:Escherichia coli; D017245:Foscarnet / Q000737:chemistry / Q000494:pharmacology; D054303:HIV Reverse Transcriptase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000187:drug effects; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D019829:Nevirapine / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid
D003848:Deoxyguanine Nucleotides; D054306:Dideoxynucleotides; D011993:Recombinant Fusion Proteins; D018894:Reverse Transcriptase Inhibitors; D017245:Foscarnet; C031709:2',3'-dideoxyguanosine 5'-triphosphate; D019829:Nevirapine; D054303:HIV Reverse Transcriptase