Herpes simplex virus thymidine kinase mutations associated with resistance to acyclovir: a site-directed mutagenesis study.
Mutations in the thymidine kinase (TK) gene of herpes simplex virus (HSV) may confer resistance to acyclovir (ACV). Because of the high genetic polymorphism of this gene, discriminating between mutations related to resistance and mutations related to gene polymorphism can be difficult, especially when no sensitive strain has been previously isolated from the same patient. To assess the role of the mutations located at codons 51, 77, 83, and 175, previously detected in HSV-1 clinical isolates (F. Morfin, G. Souillet, K. Bilger, T. Ooka, M. Aymard, and D. Thouvenot, J. Infect. Dis. 182:290-293, 2000), in the acquisition of resistance to ACV, four mutants with site-directed mutations at these respective codons were constructed. The enzymatic activity of the proteins, produced using both a reticulocyte lysate system and a bacterial system, was evaluated using [(3)H]thymidine as substrate. This site-directed mutagenesis revealed that mutations at codons 51, 83, and 175 induce a loss of HSV-1 TK activity and are thus clearly involved in the acquisition of resistance to ACV. On the other hand, the mutation at codon 77 does not affect enzyme activity.
Frobert
E
E
Laboratoire de Virologie, Hospices Civils de Lyon, Domaine Rockefeller, 8 avenue Rockefeller, 69373 Lyon cedex 08, France. e.frobert@wanadoo.fr
Ooka
T
T
Cortay
J C
JC
Lina
B
B
Thouvenot
D
D
Morfin
F
F
D000212:Acyclovir / Q000494:pharmacology*; D000998:Antiviral Agents / Q000494:pharmacology*; D024882:Drug Resistance, Viral; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D018139:Simplexvirus / Q000187:drug effects* / Q000201:enzymology*; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics*
D000998:Antiviral Agents; D013937:Thymidine Kinase; D000212:Acyclovir
J Infect Dis. 1999 Aug;180(2):487-90
10395866
J Gen Virol. 1998 Sep;79 ( Pt 9):2083-92
9747715
J Clin Microbiol. 2000 May;38(5):1839-44
10790110
J Infect Dis. 2000 Jun;181(6):2055-8
10837192
J Infect Dis. 2000 Jul;182(1):290-3
10882609
Exp Eye Res. 2000 Oct;71(4):353-9
10995556
J Virol. 2001 Apr;75(7):3105-10
11238837
Drug Resist Updat. 2002 Apr;5(2):88-114
12135584
J Clin Microbiol. 2004 Jan;42(1):242-9
14715760
J Gen Virol. 1983 Mar;64 Pt 3:523-32
6298344
Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074-8
3156376
J Gen Virol. 1986 Apr;67 ( Pt 4):753-8
3007662
J Gen Virol. 1989 Apr;70 ( Pt 4):869-79
2543763
Ann Intern Med. 1990 Mar 15;112(6):416-22
2155570
J Gen Virol. 1990 Dec;71 ( Pt 12):2979-87
2273392
J Virol. 1997 May;71(5):3872-8
9094663
Electrophoresis. 1997 May;18(5):819-25
9194614
Antimicrob Agents Chemother. 1998 Apr;42(4):868-72
9559798
J Infect Dis. 1998 Aug;178(2):297-303
9697707
J Med Virol. 1998 Oct;56(2):151-8
9746072
Antimicrob Agents Chemother. 1999 Oct;43(10):2412-6
10508017