Effect of sequence polymorphism and drug resistance on two HIV‐1 Gag processing sites

• Published in: European journal of biochemistry Vol. 269; no. 16; pp. 4114 - 4120
• Main Authors: Fehér, Anita, Weber, Irene T., Bagossi, Péter, Boross, Péter, Mahalingam, Bhuvaneshwari, Louis, John M., Copeland, Terry D., Torshin, Ivan Y., Harrison, Robert W., Tözsér, József
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01.08.2002
• Subjects: Amino Acid Sequence; Amino Acid Substitution; Binding Sites; Carbamates; Catalysis; Drug Resistance, Viral - genetics; Gag processing sites; Gene Products, gag - metabolism; Genes, gag; HIV Protease - chemistry; HIV Protease - genetics; HIV Protease - metabolism; HIV Protease Inhibitors - metabolism; HIV Protease Inhibitors - pharmacology; HIV-1 - drug effects; HIV-1 - genetics; HIV-1 - metabolism; HIV‐1 proteinase; Kinetics; Models, Molecular; molecular modeling; Molecular Sequence Data; oligopeptide substrates; Oligopeptides - metabolism; Polymorphism, Genetic; Protein Conformation; Substrate Specificity; Sulfonamides - pharmacology; Urea - analogs & derivatives; Urea - pharmacology
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1046/j.1432-1033.2002.03105.x

The HIV‐1 proteinase (PR) has proved to be a good target for antiretroviral therapy of AIDS, and various PR inhibitors are now in clinical use. However, there is a rapid selection of viral variants bearing mutations in the proteinase that are resistant to clinical inhibitors. Drug resistance also involves mutations of the nucleocapsid/p1 and p1/p6 cleavage sites of Gag, both in vitro and in vivo. Cleavages at these sites have been shown to be rate limiting steps for polyprotein processing and viral maturation. Furthermore, these sites show significant sequence polymorphism, which also may have an impact on virion infectivity. We have studied the hydrolysis of oligopeptides representing these cleavage sites with representative mutations found as natural variations or that arise as resistant mutations. Wild‐type and five drug resistant PRs with mutations within or outside the substrate binding site were tested. While the natural variations showed either increased or decreased susceptibility of peptides toward the proteinases, the resistant mutations always had a beneficial effect on catalytic efficiency. Comparison of the specificity changes obtained for the various substrates suggested that the maximization of the van der Waals contacts between substrate and PR is the major determinant of specificity: the same effect is crucial for inhibitor potency. The natural nucleocapsid/p1 and p1/p6 sites do not appear to be optimized for rapid hydrolysis. Hence, mutation of these rate limiting cleavage sites can partly compensate for the reduced catalytic activity of drug resistant mutant HIV‐1 proteinases.