Effects of the Δ67 complex of mutations in human immunodeficiency virus type 1 reverse transcriptase on nucleoside analog excision

被引:30
作者
Boyer, PL
Imamichi, T
Sarafianos, SG
Arnold, E
Hughes, SH
机构
[1] NCI, FCRDC, HIV Drug Resistance Program, Frederick, MD 21702 USA
[2] NCI, SAIC Frederick, Frederick, MD 21702 USA
[3] Rutgers State Univ, Ctr Adv Biotechnol & Med, Piscataway, NJ 08854 USA
[4] Rutgers State Univ, Dept Chem, Piscataway, NJ USA
关键词
D O I
10.1128/JVI.78.18.9987-9997.2004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Long-term use of combination therapy against human immunodeficiency virus type (HIV-1) provides strong selective pressure on the virus, and HIV-1 variants that are resistant to multiple inhibitors have been isolated. HIV-1 variants containing amino acid substitutions within the coding region of HIV-1 reverse transcriptase (RT), such as the 3'-azido-3'-deoxythymidine (AZT)-resistant variant AZT-R (M41L/D67N/K70R/T215Y/K219Q) and a variant containing an insertion in the fingers domain (S(69)SGR(70)/T215Y), are resistant to the nucleoside RT inhibitor (NRTI) AZT because of an increase in the level of excision of AZT monophosphate (AZTMP) from the primer. While rare, variants have also been isolated which contain deletions in the RT coding region. One such virus, described by Imamichi et al. (J. Virol 74:10958-10964, 2000; J. Virol. 74:10231028, 2000; J. Virol. 75:3988-3992, 2001), contains numerous amino acid substitutions and a deletion of codon 67, which we have designated the Delta67 complex of mutations. We have expressed and purified HIV-1 RT containing these mutations. We compared the polymerase and pyrophosphorolysis (excision) activity of an RT with the Delta67 complex of mutations to wild-type RT and the two other AZT-resistant variants described above. All of the AZT-resistant variants we tested excise AZTMP and 9- [2-(R)-(phosphonomethoxy)propyl] adenine (PMPA [tenofovir]) from the end of a primer more efficiently than wild-type RT. Although the variant RTs excised d4TMP less efficiently than AZTMP and PMPA, they were able to excise d4TMP more efficiently than wild-type RT. HIV-1 RT containing the Delta67 complex of mutations was not able to excise as broad a range of NRTIs as the fingers insertion variant SSGR/T215Y, but it was able to polymerize efficiently with low concentrations of deoxynucleoside triphosphates and seems to be able to excise AZTMP and PMPA at lower ATP concentrations than AZT-R or SSGR/T215Y, suggesting that a virus containing the Delta67 complex of mutations would replicate reasonably well in quiescent cells, even in the presence of AZT.
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页码:9987 / 9997
页数:11
相关论文
共 38 条
[31]  
Ross L, 2000, J HUMAN VIROL, V3, P144
[32]   Thymidine analog and multinucleoside resistance mutations are associated with decreased phenotypic susceptibility to stavudine in HIV type 1 isolated from zidovudine-naive patients experiencing viremia on stavudine-containing regimens [J].
Ross, L ;
Scarsella, A ;
Raffanti, S ;
Henry, K ;
Becker, S ;
Fisher, R ;
Liao, QM ;
Hirani, A ;
Graham, N ;
St Clair, M ;
Hernandez, J .
AIDS RESEARCH AND HUMAN RETROVIRUSES, 2001, 17 (12) :1107-1115
[33]   Structures of HIV-1 reverse transcriptase with pre- and post-translocation AZTMP-terminated DNA [J].
Sarafianos, SG ;
Clark, AD ;
Das, K ;
Tuske, S ;
Birktoft, JJ ;
Ilankumaran, P ;
Ramesha, AR ;
Sayer, JM ;
Jerina, DM ;
Boyer, PL ;
Hughes, SH ;
Arnold, E .
EMBO JOURNAL, 2002, 21 (23) :6614-6624
[34]  
Schinazi R, 2000, INT ANTIVIRAL NEWS, V8, P65
[35]   Subtle decreases in stavudine phenotypic susceptibility predict poor virologic response to stavudine monotherapy in zidovudine-experienced patients [J].
Shulman, NS ;
Hughes, MD ;
Winters, MA ;
Shafer, RW ;
Zolopa, AR ;
Hellmann, NS ;
Bates, M ;
Whitcomb, JM ;
Katzenstein, DA .
JAIDS-JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES, 2002, 31 (02) :121-127
[36]   Nucleotide-induced stable complex formation by HIV-1 reverse transcriptase [J].
Tong, WJ ;
Lu, CD ;
Sharma, SK ;
Matsuura, S ;
So, AG ;
Scott, WA .
BIOCHEMISTRY, 1997, 36 (19) :5749-5757
[37]   Structures of HIV-1 RT-DNA complexes before and after incorporation of the anti-AIDS drug tenofovir [J].
Tuske, S ;
Sarafianos, SG ;
Clark, AD ;
Ding, JP ;
Naeger, LK ;
White, KL ;
Miller, MD ;
Gibbs, CS ;
Boyer, PL ;
Clark, P ;
Wang, G ;
Gaffney, BL ;
Jones, RA ;
Jerina, DM ;
Hughes, SH ;
Arnold, E .
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2004, 11 (05) :469-474
[38]   Molecular mechanisms of tenofovir resistance conferred by human immunodeficiency virus type 1 reverse transcriptase containing a diserine insertion, after residue 69 and multiple thymidine analog-associated mutations [J].
White, KL ;
Chen, JM ;
Margot, NA ;
Wrin, T ;
Petropoulos, CJ ;
Naeger, LK ;
Swaminathan, S ;
Miller, MD .
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 2004, 48 (03) :992-1003