Inhibitors of human immunodeficiency virus type 1 zinc fingers prevent normal processing of gag precursors and result in the release of noninfectious virus particles.
The Cys-Xaa2-Cys-Xaa4-His-Xaa4-Cys zinc fingers of retroviral nucleocapsid proteins are prime antiviral targets because of conservation of the Cys and His chelating residues and the absolute requirement of these fingers in both early and late phases of retroviral replication. We previously reported that certain disulfide-substituted benzamides (DIBAs) chemically modify the Cys residues of the fingers, resulting in inhibition of human immunodeficiency virus type 1 (HIV-1) replication (W. G. Rice, J. G. Supko, L. Malspeis, R. W. Buckheit, Jr., D. Clanton, M. Bu, L. Graham, C. A. Schaeffer, J. A. Turpin, J. Domagala, R. Gogliotti, J. P. Bader, S. M. Halliday, L. Coren, R. C. Sowder II, L. O. Arthur, and L. E. Henderson, Science 270:1194-1197, 1995). We now examine the consequences of the interaction of DIBAs with the zinc fingers of the HIV-1 p7 nucleocapsid protein and its Pr55gag precursor. In HIV-1-infected U1 cells, DIBAs inhibited the release of infectious virions, and even under conditions in which virion particles were produced, the particles were noninfectious. DIBAs caused abnormal processing of Gag precursors, and the inhibitory effect on processing was not due to inhibition of the HIV-1 protease enzyme or Pr55gag myristoylation. Rather, the defect in processing was due to the formation of intermolecular cross-linkages among the zinc fingers of adjacent Gag molecules, rendering the precursors no longer recognizable by HIV-1 protease. Likewise, DIBAs caused intermolecular cross-linkage among recombinant Pr55gag packaged into pseudovirions, thereby generating modified precursors that were resistant to the action of protease. Thus, DIBAs chemically modified the mutationally intolerant retroviral zinc fingers in infected cells, interrupting protease-mediated maturation of virions and leading ultimately to the production of compromised virions.
Turpin
J A
JA
Laboratory of Antiviral Drug Mechanisms, National Cancer Institute-Frederick Cancer Research and Development Center, SAIC Frederick, Maryland 21702, USA.
Terpening
S J
SJ
Schaeffer
C A
CA
Yu
G
G
Glover
C J
CJ
Felsted
R L
RL
Sausville
E A
EA
Rice
W G
WG
D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D001549:Benzamides / Q000494:pharmacology*; D015153:Blotting, Western; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000494:pharmacology; D015683:Gene Products, gag / Q000096:biosynthesis* / Q000378:metabolism*; D016333:HIV Protease / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000096:biosynthesis / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014758:Viral Core Proteins / Q000737:chemistry / Q000378:metabolism*; D016335:Zinc Fingers / Q000187:drug effects
D000998:Antiviral Agents; D001549:Benzamides; D003432:Cross-Linking Reagents; C100805:DIBA-1; C100806:DIBA-3; D015683:Gene Products, gag; D011498:Protein Precursors; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D014758:Viral Core Proteins; C059767:p55 gag precursor protein, Human immunodeficiency virus 1; D016333:HIV Protease