Impact of novel human immunodeficiency virus type 1 reverse transcriptase mutations P119S and T165A on 4'-ethynylthymidine analog resistance profile.
2',3'-Didehydro-3'-deoxy-4'-ethynylthymidine (4'-Ed4T), a derivative of stavudine (d4T), has potent activity against human immunodeficiency virus and is much less inhibitory to mitochondrial DNA synthesis and cell growth than its progenitor, d4T. 4'-Ed4T triphosphate was a better reverse transcriptase (RT) inhibitor than d4T triphosphate, due to the additional binding of the 4'-ethynyl group at a presumed hydrophobic pocket in the RT active site. Previous in vitro selection for 4'-Ed4T-resistant viral strains revealed M184V and P119S/T165A/M184V mutations on days 26 and 81, respectively; M184V and P119S/T165A/M184V conferred 3- and 130-fold resistance to 4'-Ed4T, respectively. We investigated the relative contributions of these mutations, engineered into the strain NL4-3 background, to drug resistance, RT activity, and viral growth. Viral variants with single RT mutations (P119S or T165A) did not show resistance to 4'-Ed4T; however, M184V and P119S/T165A/M184V conferred three- and fivefold resistance, respectively, compared with that of the wild-type virus. The P119S/M184V and T165A/M184V variants showed about fourfold resistance to 4'-Ed4T. The differences in the growth kinetics of the variants were not more than threefold. The purified RT of mutants with the P119S/M184V and T165A/M184V mutations were inhibited by 4'-Ed4TTP with 8- to 13-fold less efficiency than wild-type RT. M184V may be the primary resistance-associated mutation of 4'-Ed4T, and P119S and T165A are secondary mutations. On the basis of our findings and the results of structural modeling, a virus with a high degree of resistance to 4'-Ed4T (e.g., more than 50-fold resistance) will be difficult to develop. The previously observed 130-fold resistance of the virus with P119S/T165A/M184V to 4'-Ed4T may be partly due to mutations both in the RT sequence and outside the RT sequence.
Yang
Guangwei
G
Department of Pharmacology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Paintsil
Elijah
E
Dutschman
Ginger E
GE
Grill
Susan P
SP
Wang
Chuan-Jen
CJ
Wang
Jimin
J
Tanaka
Hiromichi
H
Hamasaki
Takayuki
T
Baba
Masanori
M
Cheng
Yung-Chi
YC
D019380:Anti-HIV Agents / Q000494:pharmacology*; D003198:Computer Simulation; D024882:Drug Resistance, Viral; D054303:HIV Reverse Transcriptase / Q000235:genetics*; D015497:HIV-1 / Q000187:drug effects*; D006801:Humans; D008826:Microbial Sensitivity Tests; D009154:Mutation; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D018119:Stavudine / Q000031:analogs & derivatives* / Q000494:pharmacology
D019380:Anti-HIV Agents; D018894:Reverse Transcriptase Inhibitors; C486840:2',3'-didehydro-3'-deoxy-4'-ethynylthymidine; D018119:Stavudine; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase
Biochem Biophys Res Commun. 1987 Jan 15;142(1):128-34
3028398
EMBO J. 1996 Aug 1;15(15):4040-9
8670908
Antimicrob Agents Chemother. 2005 Aug;49(8):3355-60
16048947
Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10027-32
10468556
Antimicrob Agents Chemother. 1987 Jun;31(6):907-10
3039911
Antivir Ther. 2001;6 Suppl 3:1-14
11678469
AIDS. 2002 Dec 6;16(18):2391-8
12461412
Antimicrob Agents Chemother. 2004 May;48(5):1640-6
15105115
J Virol. 1991 Oct;65(10):5232-6
1716689
Virology. 2004 Apr 25;322(1):13-21
15063112
J Biol Chem. 2002 Aug 30;277(35):31593-600
12080078
Antimicrob Agents Chemother. 1994 Dec;38(12):2743-9
7695256
J Infect Dis. 1991 Dec;164(6):1075-81
1720152
J Biol Chem. 2001 Nov 2;276(44):40847-57
11526116
Bioorg Med Chem Lett. 2003 Nov 3;13(21):3775-7
14552777
J Virol. 1988 Dec;62(12):4691-6
2846880
Science. 1998 Nov 27;282(5394):1669-75
9831551
N Engl J Med. 2004 Mar 4;350(10):1023-35
14999114
Antimicrob Agents Chemother. 2004 Apr;48(4):1300-6
15047533
Antimicrob Agents Chemother. 2002 Jun;46(6):1896-905
12019106
AIDS. 2007 Mar 30;21(6):665-75
17413687
J Med Chem. 2004 Oct 7;47(21):5041-8
15456247
Virology. 2003 Jun 20;311(1):202-12
12832217
Science. 1995 Aug 4;269(5224):696-9
7542804
Int J Biochem Cell Biol. 2008;40(11):2410-20
18487070
Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1934-8
1371886
J Virol. 1994 Mar;68(3):1660-6
7509000
J Virol. 1999 Oct;73(10):8448-56
10482597
J Virol. 1986 Aug;59(2):284-91
3016298
Science. 1996 Mar 1;271(5253):1282-5
8638110
J Virol. 2002 Oct;76(19):9600-13
12208939
Antiviral Res. 2007 Mar;73(3):185-91
17109975
AIDS Res Ther. 2009 Apr 22;6:5
19386130
Science. 1995 Jan 27;267(5197):483-9
7824947
Antimicrob Agents Chemother. 2009 Aug;53(8):3317-24
19470503
AIDS. 1991 Feb;5(2):137-44
1709563
Antimicrob Agents Chemother. 2008 Jun;52(6):2035-42
18391035
AIDS. 1998 Oct 1;12(14):1735-44
9792373
J Virol. 2004 Jul;78(14):7545-52
15220429
Antiviral Res. 2006 Sep;71(2-3):322-34
16716415
Antimicrob Agents Chemother. 2002 Jan;46(1):89-94
11751116
Antimicrob Agents Chemother. 1993 Jun;37(6):1390-2
8392313
J Virol. 2004 Jan;78(2):603-11
14694092
Clin Ther. 2000 Jun;22(6):685-708
10929917
Antimicrob Agents Chemother. 1994 Aug;38(8):1824-8
7986014
Biochemistry. 1999 Jul 20;38(29):9440-8
10413520
Biochem Pharmacol. 1987 Sep 1;36(17):2713-8
2443141
J Infect Dis. 1992 Jan;165(1):105-10
1370174
Science. 1989 Dec 1;246(4934):1155-8
2479983
J Infect Dis. 1995 Jun;171(6):1411-9
7539472