Search tips
Search criteria 


Logo of aacPermissionsJournals.ASM.orgJournalAAC ArticleJournal InfoAuthorsReviewers
Antimicrob Agents Chemother. 1996 August; 40(8): 1957–1960.
PMCID: PMC163451

Generation of duck hepatitis B virus polymerase mutants through site-directed mutagenesis which demonstrate resistance to lamivudine [(--)-beta-L-2', 3'-dideoxy-3'-thiacytidine] in vitro.


Hepatitis B virus replication is very sensitive to lamivudine. A single amino acid change in human immunodeficiency virus reverse transcriptase is responsible for high-level resistance to this compound. Duck hepatitis B virus mutants were created bearing the analogous amino acid change in the duck hepatitis B virus polymerase. Viral DNA production was reduced 92% for the wild-type virus at 2 micrograms of lamivudine per ml, while the mutants required 40 micrograms of lamivudine per ml to inhibit replication by greater than 80%.

Full Text

The Full Text of this article is available as a PDF (246K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Benhamou Y, Dohin E, Lunel-Fabiani F, Poynard T, Huraux JM, Katlama C, Opolon P, Gentilini M. Efficacy of lamivudine on replication of hepatitis B virus in HIV-infected patients. Lancet. 1995 Feb 11;345(8946):396–397. [PubMed]
  • Boucher CA, Cammack N, Schipper P, Schuurman R, Rouse P, Wainberg MA, Cameron JM. High-level resistance to (-) enantiomeric 2'-deoxy-3'-thiacytidine in vitro is due to one amino acid substitution in the catalytic site of human immunodeficiency virus type 1 reverse transcriptase. Antimicrob Agents Chemother. 1993 Oct;37(10):2231–2234. [PMC free article] [PubMed]
  • Chang CN, Doong SL, Zhou JH, Beach JW, Jeong LS, Chu CK, Tsai CH, Cheng YC, Liotta D, Schinazi R. Deoxycytidine deaminase-resistant stereoisomer is the active form of (+/-)-2',3'-dideoxy-3'-thiacytidine in the inhibition of hepatitis B virus replication. J Biol Chem. 1992 Jul 15;267(20):13938–13942. [PubMed]
  • Chang CN, Skalski V, Zhou JH, Cheng YC. Biochemical pharmacology of (+)- and (-)-2',3'-dideoxy-3'-thiacytidine as anti-hepatitis B virus agents. J Biol Chem. 1992 Nov 5;267(31):22414–22420. [PubMed]
  • Coates JA, Cammack N, Jenkinson HJ, Jowett AJ, Jowett MI, Pearson BA, Penn CR, Rouse PL, Viner KC, Cameron JM. (-)-2'-deoxy-3'-thiacytidine is a potent, highly selective inhibitor of human immunodeficiency virus type 1 and type 2 replication in vitro. Antimicrob Agents Chemother. 1992 Apr;36(4):733–739. [PMC free article] [PubMed]
  • Coates JA, Cammack N, Jenkinson HJ, Mutton IM, Pearson BA, Storer R, Cameron JM, Penn CR. The separated enantiomers of 2'-deoxy-3'-thiacytidine (BCH 189) both inhibit human immunodeficiency virus replication in vitro. Antimicrob Agents Chemother. 1992 Jan;36(1):202–205. [PMC free article] [PubMed]
  • Condreay LD, Aldrich CE, Coates L, Mason WS, Wu TT. Efficient duck hepatitis B virus production by an avian liver tumor cell line. J Virol. 1990 Jul;64(7):3249–3258. [PMC free article] [PubMed]
  • Condreay LD, Jansen RW, Powdrill TF, Johnson LC, Selleseth DW, Paff MT, Daluge SM, Painter GR, Furman PA, Ellis MN, et al. Evaluation of the potent anti-hepatitis B virus agent (-) cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine in a novel in vivo model. Antimicrob Agents Chemother. 1994 Mar;38(3):616–619. [PMC free article] [PubMed]
  • Doong SL, Tsai CH, Schinazi RF, Liotta DC, Cheng YC. Inhibition of the replication of hepatitis B virus in vitro by 2',3'-dideoxy-3'-thiacytidine and related analogues. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8495–8499. [PubMed]
  • Gao Q, Gu Z, Hiscott J, Dionne G, Wainberg MA. Generation of drug-resistant variants of human immunodeficiency virus type 1 by in vitro passage in increasing concentrations of 2',3'-dideoxycytidine and 2',3'-dideoxy-3'-thiacytidine. Antimicrob Agents Chemother. 1993 Jan;37(1):130–133. [PMC free article] [PubMed]
  • Girones R, Miller RH. Mutation rate of the hepadnavirus genome. Virology. 1989 Jun;170(2):595–597. [PubMed]
  • Gosselin G, Schinazi RF, Sommadossi JP, Mathé C, Bergogne MC, Aubertin AM, Kirn A, Imbach JL. Anti-human immunodeficiency virus activities of the beta-L enantiomer of 2',3'-dideoxycytidine and its 5-fluoro derivative in vitro. Antimicrob Agents Chemother. 1994 Jun;38(6):1292–1297. [PMC free article] [PubMed]
  • Hart GJ, Orr DC, Penn CR, Figueiredo HT, Gray NM, Boehme RE, Cameron JM. Effects of (-)-2'-deoxy-3'-thiacytidine (3TC) 5'-triphosphate on human immunodeficiency virus reverse transcriptase and mammalian DNA polymerases alpha, beta, and gamma. Antimicrob Agents Chemother. 1992 Aug;36(8):1688–1694. [PMC free article] [PubMed]
  • Larder BA, Kemp SD, Harrigan PR. Potential mechanism for sustained antiretroviral efficacy of AZT-3TC combination therapy. Science. 1995 Aug 4;269(5224):696–699. [PubMed]
  • Lin TS, Luo MZ, Liu MC, Pai SB, Dutschman GE, Cheng YC. Antiviral activity of 2',3'-dideoxy-beta-L-5-fluorocytidine (beta-L-FddC) and 2',3'-dideoxy-beta-L-cytidine (beta-L-ddC) against hepatitis B virus and human immunodeficiency virus type 1 in vitro. Biochem Pharmacol. 1994 Jan 20;47(2):171–174. [PubMed]
  • Lin TS, Luo MZ, Liu MC, Pai SB, Dutschman GE, Cheng YC. Synthesis and biological evaluation of 2',3'-dideoxy-L-pyrimidine nucleosides as potential antiviral agents against human immunodeficiency virus (HIV) and hepatitis B virus (HBV). J Med Chem. 1994 Mar 18;37(6):798–803. [PubMed]
  • Mandart E, Kay A, Galibert F. Nucleotide sequence of a cloned duck hepatitis B virus genome: comparison with woodchuck and human hepatitis B virus sequences. J Virol. 1984 Mar;49(3):782–792. [PMC free article] [PubMed]
  • Paff MT, Averett DR, Prus KL, Miller WH, Nelson DJ. Intracellular metabolism of (-)- and (+)-cis-5-fluoro-1-[2-(hydroxymethyl)-1,3-oxathiolan-5-yl]cytosine in HepG2 derivative 2.2.15 (subclone P5A) cells. Antimicrob Agents Chemother. 1994 Jun;38(6):1230–1238. [PMC free article] [PubMed]
  • Schalm SW, de Man RA, Heijtink RA, Niesters HG. New nucleoside analogues for chronic hepatitis B. J Hepatol. 1995;22(1 Suppl):52–56. [PubMed]
  • Schinazi RF, Gosselin G, Faraj A, Korba BE, Liotta DC, Chu CK, Mathé C, Imbach JL, Sommadossi JP. Pure nucleoside enantiomers of beta-2',3'-dideoxycytidine analogs are selective inhibitors of hepatitis B virus in vitro. Antimicrob Agents Chemother. 1994 Sep;38(9):2172–2174. [PMC free article] [PubMed]
  • Schinazi RF, Lloyd RM, Jr, Nguyen MH, Cannon DL, McMillan A, Ilksoy N, Chu CK, Liotta DC, Bazmi HZ, Mellors JW. Characterization of human immunodeficiency viruses resistant to oxathiolane-cytosine nucleosides. Antimicrob Agents Chemother. 1993 Apr;37(4):875–881. [PMC free article] [PubMed]
  • Severini A, Liu XY, Wilson JS, Tyrrell DL. Mechanism of inhibition of duck hepatitis B virus polymerase by (-)-beta-L-2',3'-dideoxy-3'-thiacytidine. Antimicrob Agents Chemother. 1995 Jul;39(7):1430–1435. [PMC free article] [PubMed]
  • Skalski V, Chang CN, Dutschman G, Cheng YC. The biochemical basis for the differential anti-human immunodeficiency virus activity of two cis enantiomers of 2',3'-dideoxy-3'-thiacytidine. J Biol Chem. 1993 Nov 5;268(31):23234–23238. [PubMed]
  • Staprans S, Loeb DD, Ganem D. Mutations affecting hepadnavirus plus-strand DNA synthesis dissociate primer cleavage from translocation and reveal the origin of linear viral DNA. J Virol. 1991 Mar;65(3):1255–1262. [PMC free article] [PubMed]
  • Sullivan V, Biron KK, Talarico C, Stanat SC, Davis M, Pozzi LM, Coen DM. A point mutation in the human cytomegalovirus DNA polymerase gene confers resistance to ganciclovir and phosphonylmethoxyalkyl derivatives. Antimicrob Agents Chemother. 1993 Jan;37(1):19–25. [PMC free article] [PubMed]
  • Tisdale M, Kemp SD, Parry NR, Larder BA. Rapid in vitro selection of human immunodeficiency virus type 1 resistant to 3'-thiacytidine inhibitors due to a mutation in the YMDD region of reverse transcriptase. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5653–5656. [PubMed]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)