Analysis of HIV-2 RT mutants provides evidence that resistance of HIV-1 RT and HIV-2 RT to nucleoside analogs involves a repositioning of the template-primer.
Mutations that confer resistance to nucleoside analogs do not cluster around the deoxynucleotide triphosphate (dNTP) binding site. Instead, these mutations appear to lie along the groove in the enzyme where the template-primer binds. Based on such structural data and on complementary biochemical analyses, it has been suggested that resistance to nucleoside analogs involves repositioning of the template-primer. We have prepared mutations in HIV-2 RT that are the homologs of mutations that confer resistance to nucleoside analogs in HIV-1 RT. Analysis of the behavior of HIV-2 RT mutants (Leu74Val, Glu89Gly, Ser215Tyr, Leu74Val/Ser215Tyr and Glu89Gly/Ser215Tyr) in vitro confirms the results obtained with HIV-1 RT: resistance is a function of the length of the template overhang. These analyses also suggest that the homolog in HIV-2 RT of one of the mutations that confers resistance to AZT in HIV-1 RT (Thr215Tyr) confers resistance by repositioning of the template-primer.
Perach
M
M
Department of Cell Biology and Histology, Sackler School of Medicine, Tel Aviv University, Israel.
Rubinek
T
T
Hughes
S H
SH
Hizi
A
A
D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry*; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000378:metabolism*; D054306:Dideoxynucleotides; D004352:Drug Resistance, Microbial / Q000235:genetics; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology*; D015498:HIV-2 / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D008958:Models, Molecular; D009154:Mutation; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors / Q000378:metabolism*; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000378:metabolism
D017931:DNA Primers; D004279:DNA, Viral; D003848:Deoxyguanine Nucleotides; D003854:Deoxyribonucleotides; D054306:Dideoxynucleotides; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; C031709:2',3'-dideoxyguanosine 5'-triphosphate; C029603:deoxyguanosine triphosphate; C072416:reverse transcriptase, Human immunodeficiency virus 2; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; C017216:2',3'-dideoxythymidine triphosphate; C024157:thymidine 5'-triphosphate