Relationship between 3'-azido-3'-deoxythymidine resistance and primer unblocking activity in foscarnet-resistant mutants of human immunodeficiency virus type 1 reverse transcriptase.
Phosphonoformate (foscarnet) is a pyrophosphate (PP(i)) analogue and a potent inhibitor of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT), acting through the PP(i) binding site on the enzyme. HIV-1 RT can unblock a chain-terminated DNA primer by phosphorolytic transfer of the terminal residue to an acceptor substrate (PP(i) or a nucleotide such as ATP) which also interacts with the PP(i) binding site. Primer-unblocking activity is increased in mutants of HIV-1 that are resistant to the chain-terminating nucleoside inhibitor 3'-azido-3'-deoxythymidine (AZT). We have compared the primer-unblocking activity for HIV-1 RT containing various foscarnet resistance mutations (K65R, W88G, W88S, E89K, S117T, Q161L, M164I, and the double mutant Q161L/H208Y) alone or in combination with AZT resistance mutations. The level of primer-unblocking activity varied over a 150-fold range for these enzymes and was inversely correlated with foscarnet resistance and directly correlated with AZT resistance. Based on published crystal structures of HIV-1 RT, many of the foscarnet resistance mutations affect residues that do not make direct contact with the catalytic residues of RT, the incoming deoxynucleoside triphosphate (dNTP), or the primer-template. These mutations may confer foscarnet resistance and reduce primer unblocking by indirectly decreasing the binding and retention of foscarnet, PP(i), and ATP. Alternatively, the binding position or orientation of PP(i), ATP, or the primer-template may be changed in the mutant enzyme complex so that molecular interactions required for the unblocking reaction are impaired while dNTP binding and incorporation are not.
Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Florida 33101, USA.
Peter R
PR
Meyer
Suzanne E
SE
Matsuura
Dianna
D
Zonarich
Rahul R
RR
Chopra
Eric
E
Pendarvis
Holly Z
HZ
Bazmi
John W
JW
Mellors
Walter A
WA
Scott
D000998:Antiviral Agents / Q000494:pharmacology*; D001665:Binding Sites; D002460:Cell Line; D017931:DNA Primers; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D054306:Dideoxynucleotides; D011756:Diphosphates / Q000378:metabolism; D024882:Drug Resistance, Viral / Q000235:genetics; D017245:Foscarnet / Q000494:pharmacology*; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D009154:Mutation; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000378:metabolism; D015215:Zidovudine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology
D000998:Antiviral Agents; D017931:DNA Primers; D003838:Deoxyadenine Nucleotides; D054306:Dideoxynucleotides; D011756:Diphosphates; D013942:Thymine Nucleotides; C010110:2',3'-dideoxyadenosine 5'-phosphate; C057287:3'-azido-3'-deoxythymidine 5'phosphate; D017245:Foscarnet; D015215:Zidovudine; D054303:HIV Reverse Transcriptase