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J Virol. Feb 1997; 71(2): 1089–1096.
PMCID: PMC191160

Impaired fitness of human immunodeficiency virus type 1 variants with high-level resistance to protease inhibitors.


One hope to maintain the benefits of antiviral therapy against the human immunodeficiency virus type 1 (HIV-1), despite the development of resistance, is the possibility that resistant variants will show decreased viral fitness. To study this possibility, HIV-1 variants showing high-level resistance (up to 1,500-fold) to the substrate analog protease inhibitors BILA 1906 BS and BILA 2185 BS have been characterized. Active-site mutations V32I and I84V/A were consistently observed in the protease of highly resistant viruses, along with up to six other mutations. In vitro studies with recombinant mutant proteases demonstrated that these mutations resulted in up to 10(4)-fold increases in the Ki values toward BILA 1906 BS and BILA 2185 BS and a concomitant 2,200-fold decrease in catalytic efficiency of the enzymes toward a synthetic substrate. When introduced into viral molecular clones, the protease mutations impaired polyprotein processing, consistent with a decrease in enzyme activity in virions. Despite these observations, however, most mutations had little effect on viral replication except when the active-site mutations V32I and I84V/A were coexpressed in the protease. The latter combinations not only conferred a significant growth reduction of viral clones on peripheral blood mononuclear cells but also caused the complete disappearance of mutated clones when cocultured with wild-type virus on T-cell lines. Furthermore, the double nucleotide mutation I84A rapidly reverted to I84V upon drug removal, confirming its impact on viral fitness. Therefore, high-level resistance to protease inhibitors can be associated with impaired viral fitness, suggesting that antiviral therapies with such inhibitors may maintain some clinical benefits.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adachi A, Gendelman HE, Koenig S, Folks T, Willey R, Rabson A, Martin MA. Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. J Virol. 1986 Aug;59(2):284–291. [PMC free article] [PubMed]
  • Condra JH, Schleif WA, Blahy OM, Gabryelski LJ, Graham DJ, Quintero JC, Rhodes A, Robbins HL, Roth E, Shivaprakash M, et al. In vivo emergence of HIV-1 variants resistant to multiple protease inhibitors. Nature. 1995 Apr 6;374(6522):569–571. [PubMed]
  • Darke PL, Huff JR. HIV protease as an inhibitor target for the treatment of AIDS. Adv Pharmacol. 1994;25:399–454. [PubMed]
  • Darke PL, Nutt RF, Brady SF, Garsky VM, Ciccarone TM, Leu CT, Lumma PK, Freidinger RM, Veber DF, Sigal IS. HIV-1 protease specificity of peptide cleavage is sufficient for processing of gag and pol polyproteins. Biochem Biophys Res Commun. 1988 Oct 14;156(1):297–303. [PubMed]
  • De Clercq E. HIV inhibitors targeted at the reverse transcriptase. AIDS Res Hum Retroviruses. 1992 Feb;8(2):119–134. [PubMed]
  • Doyon L, Croteau G, Thibeault D, Poulin F, Pilote L, Lamarre D. Second locus involved in human immunodeficiency virus type 1 resistance to protease inhibitors. J Virol. 1996 Jun;70(6):3763–3769. [PMC free article] [PubMed]
  • Erice A, Balfour HH., Jr Resistance of human immunodeficiency virus type 1 to antiretroviral agents: a review. Clin Infect Dis. 1994 Feb;18(2):149–156. [PubMed]
  • Erickson JW. The not-so-great escape. Nat Struct Biol. 1995 Jul;2(7):523–529. [PubMed]
  • Gulnik SV, Suvorov LI, Liu B, Yu B, Anderson B, Mitsuya H, Erickson JW. Kinetic characterization and cross-resistance patterns of HIV-1 protease mutants selected under drug pressure. Biochemistry. 1995 Jul 25;34(29):9282–9287. [PubMed]
  • Ho DD, Neumann AU, Perelson AS, Chen W, Leonard JM, Markowitz M. Rapid turnover of plasma virions and CD4 lymphocytes in HIV-1 infection. Nature. 1995 Jan 12;373(6510):123–126. [PubMed]
  • Ho DD, Toyoshima T, Mo H, Kempf DJ, Norbeck D, Chen CM, Wideburg NE, Burt SK, Erickson JW, Singh MK. Characterization of human immunodeficiency virus type 1 variants with increased resistance to a C2-symmetric protease inhibitor. J Virol. 1994 Mar;68(3):2016–2020. [PMC free article] [PubMed]
  • Kaplan AH, Michael SF, Wehbie RS, Knigge MF, Paul DA, Everitt L, Kempf DJ, Norbeck DW, Erickson JW, Swanstrom R. Selection of multiple human immunodeficiency virus type 1 variants that encode viral proteases with decreased sensitivity to an inhibitor of the viral protease. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5597–5601. [PMC free article] [PubMed]
  • Kimberlin DW, Coen DM, Biron KK, Cohen JI, Lamb RA, McKinlay M, Emini EA, Whitley RJ. Molecular mechanisms of antiviral resistance. Antiviral Res. 1995 Apr;26(4):369–401. [PubMed]
  • Kohl NE, Emini EA, Schleif WA, Davis LJ, Heimbach JC, Dixon RA, Scolnick EM, Sigal IS. Active human immunodeficiency virus protease is required for viral infectivity. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4686–4690. [PMC free article] [PubMed]
  • Larder BA, Kemp SD. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science. 1989 Dec 1;246(4934):1155–1158. [PubMed]
  • Lin Y, Lin X, Hong L, Foundling S, Heinrikson RL, Thaisrivongs S, Leelamanit W, Raterman D, Shah M, Dunn BM, et al. Effect of point mutations on the kinetics and the inhibition of human immunodeficiency virus type 1 protease: relationship to drug resistance. Biochemistry. 1995 Jan 31;34(4):1143–1152. [PubMed]
  • Lynn DE. A BASIC computer program for analyzing endpoint assays. Biotechniques. 1992 Jun;12(6):880–881. [PubMed]
  • Markowitz M, Mo H, Kempf DJ, Norbeck DW, Bhat TN, Erickson JW, Ho DD. Selection and analysis of human immunodeficiency virus type 1 variants with increased resistance to ABT-538, a novel protease inhibitor. J Virol. 1995 Feb;69(2):701–706. [PMC free article] [PubMed]
  • Molla A, Korneyeva M, Gao Q, Vasavanonda S, Schipper PJ, Mo HM, Markowitz M, Chernyavskiy T, Niu P, Lyons N, et al. Ordered accumulation of mutations in HIV protease confers resistance to ritonavir. Nat Med. 1996 Jul;2(7):760–766. [PubMed]
  • Morrison JF, Walsh CT. The behavior and significance of slow-binding enzyme inhibitors. Adv Enzymol Relat Areas Mol Biol. 1988;61:201–301. [PubMed]
  • Pargellis CA, Morelock MM, Graham ET, Kinkade P, Pav S, Lubbe K, Lamarre D, Anderson PC. Determination of kinetic rate constants for the binding of inhibitors to HIV-1 protease and for the association and dissociation of active homodimer. Biochemistry. 1994 Oct 18;33(41):12527–12534. [PubMed]
  • Partaledis JA, Yamaguchi K, Tisdale M, Blair EE, Falcione C, Maschera B, Myers RE, Pazhanisamy S, Futer O, Cullinan AB, et al. In vitro selection and characterization of human immunodeficiency virus type 1 (HIV-1) isolates with reduced sensitivity to hydroxyethylamino sulfonamide inhibitors of HIV-1 aspartyl protease. J Virol. 1995 Sep;69(9):5228–5235. [PMC free article] [PubMed]
  • Patick AK, Rose R, Greytok J, Bechtold CM, Hermsmeier MA, Chen PT, Barrish JC, Zahler R, Colonno RJ, Lin PF. Characterization of a human immunodeficiency virus type 1 variant with reduced sensitivity to an aminodiol protease inhibitor. J Virol. 1995 Apr;69(4):2148–2152. [PMC free article] [PubMed]
  • Pav S, Lubbe K, Dô F, Lamarre D, Pargellis C, Tong L. Microtube batch protein crystallization: applications to human immunodeficiency virus type 2 (HIV-2) protease and human renin. Proteins. 1994 Sep;20(1):98–102. [PubMed]
  • Peng C, Ho BK, Chang TW, Chang NT. Role of human immunodeficiency virus type 1-specific protease in core protein maturation and viral infectivity. J Virol. 1989 Jun;63(6):2550–2556. [PMC free article] [PubMed]
  • Popovic M, Read-Connole E, Gallo RC. T4 positive human neoplastic cell lines susceptible to and permissive for HTLV-III. Lancet. 1984 Dec 22;2(8417-8418):1472–1473. [PubMed]
  • Popovic M, Sarngadharan MG, Read E, Gallo RC. Detection, isolation, and continuous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science. 1984 May 4;224(4648):497–500. [PubMed]
  • Preston BD, Poiesz BJ, Loeb LA. Fidelity of HIV-1 reverse transcriptase. Science. 1988 Nov 25;242(4882):1168–1171. [PubMed]
  • Ratner L, Haseltine W, Patarca R, Livak KJ, Starcich B, Josephs SF, Doran ER, Rafalski JA, Whitehorn EA, Baumeister K, et al. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature. 1985 Jan 24;313(6000):277–284. [PubMed]
  • Rosé JR, Babé LM, Craik CS. Defining the level of human immunodeficiency virus type 1 (HIV-1) protease activity required for HIV-1 particle maturation and infectivity. J Virol. 1995 May;69(5):2751–2758. [PMC free article] [PubMed]
  • Rose RE, Gong YF, Greytok JA, Bechtold CM, Terry BJ, Robinson BS, Alam M, Colonno RJ, Lin PF. Human immunodeficiency virus type 1 viral background plays a major role in development of resistance to protease inhibitors. Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1648–1653. [PMC free article] [PubMed]
  • Sardana VV, Schlabach AJ, Graham P, Bush BL, Condra JH, Culberson JC, Gotlib L, Graham DJ, Kohl NE, LaFemina RL, et al. Human immunodeficiency virus type 1 protease inhibitors: evaluation of resistance engendered by amino acid substitutions in the enzyme's substrate binding site. Biochemistry. 1994 Mar 1;33(8):2004–2010. [PubMed]
  • Tantillo C, Ding J, Jacobo-Molina A, Nanni RG, Boyer PL, Hughes SH, Pauwels R, Andries K, Janssen PA, Arnold E. Locations of anti-AIDS drug binding sites and resistance mutations in the three-dimensional structure of HIV-1 reverse transcriptase. Implications for mechanisms of drug inhibition and resistance. J Mol Biol. 1994 Oct 28;243(3):369–387. [PubMed]
  • Tong L, Pav S, Mui S, Lamarre D, Yoakim C, Beaulieu P, Anderson PC. Crystal structures of HIV-2 protease in complex with inhibitors containing the hydroxyethylamine dipeptide isostere. Structure. 1995 Jan 15;3(1):33–40. [PubMed]
  • Toth MV, Marshall GR. A simple, continuous fluorometric assay for HIV protease. Int J Pept Protein Res. 1990 Dec;36(6):544–550. [PubMed]
  • Tözsér J, Bláha I, Copeland TD, Wondrak EM, Oroszlan S. Comparison of the HIV-1 and HIV-2 proteinases using oligopeptide substrates representing cleavage sites in Gag and Gag-Pol polyproteins. FEBS Lett. 1991 Apr 9;281(1-2):77–80. [PubMed]
  • Wei X, Ghosh SK, Taylor ME, Johnson VA, Emini EA, Deutsch P, Lifson JD, Bonhoeffer S, Nowak MA, Hahn BH, et al. Viral dynamics in human immunodeficiency virus type 1 infection. Nature. 1995 Jan 12;373(6510):117–122. [PubMed]
  • West ML, Fairlie DP. Targeting HIV-1 protease: a test of drug-design methodologies. Trends Pharmacol Sci. 1995 Feb;16(2):67–75. [PubMed]
  • Wondrak EM, Louis JM, de Rocquigny H, Chermann JC, Roques BP. The gag precursor contains a specific HIV-1 protease cleavage site between the NC (P7) and P1 proteins. FEBS Lett. 1993 Oct 25;333(1-2):21–24. [PubMed]

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