Anti-HIV agents that selectively target retroviral nucleocapsid protein zinc fingers without affecting cellular zinc finger proteins.
Agents that target the two highly conserved Zn fingers of the human immunodeficiency virus (HIV) nucleocapsid p7 (NCp7) protein are under development as antivirals. These agents covalently modify Zn-coordinating cysteine thiolates of the fingers, causing Zn ejection, loss of native protein structure and nucleic acid binding capacity, and disruption of virus replication. Concentrations of three antiviral agents that promoted in vitro Zn ejection from NCp7 and inhibited HIV replication did not impact the functions of cellular Zn finger proteins, including poly(ADP-ribose) polymerase and the Sp1 and GATA-1 transcription factors, nor did the compounds inhibit HeLa nuclear extract mediated transcription. Selectivity of interactions of these agents with NCp7 was supported by molecular modeling analysis which (1) identified a common saddle-shaped nucleophilic region on the surfaces of both NCp7 Zn fingers, (2) indicated a strong correspondence between computationally docked positions for the agents tested and overlap of frontier orbitals within the nucleophilic loci of the NCp7 Zn fingers, and (3) revealed selective steric exclusion of the agents from the core of the GATA-1 Zn finger. Further modeling analysis suggests that the thiolate of Cys49 in the carboxy-terminal finger is the site most susceptible to electrophilic attack. These data provide the first experimental evidence and rationale for antiviral agents that selectively target retroviral nucleocapsid protein Zn fingers.
Huang
M
M
Laboratory of Antiviral Drug Mechanisms, National Cancer Institute-Frederick Cancer Research and Development Center, SAIC Frederick, Frederick, Maryland 21702-1201, USA.
Maynard
A
A
Turpin
J A
JA
Graham
L
L
Janini
G M
GM
Covell
D G
DG
Rice
W G
WG
D000818:Animals; D019380:Anti-HIV Agents / Q000378:metabolism / Q000494:pharmacology*; D001391:Azo Compounds / Q000378:metabolism / Q000494:pharmacology; D001549:Benzamides / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D002213:Capsid / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D036022:Capsid Proteins; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D015683:Gene Products, gag / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D015497:HIV-1 / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D051863:Host Cell Factor C1; D006801:Humans; D008024:Ligands; D051379:Mice; D008958:Models, Molecular; D009603:Nitroso Compounds / Q000378:metabolism / Q000494:pharmacology; D050811:Octamer Transcription Factor-1; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000737:chemistry / Q000378:metabolism; D013450:Sulfones / Q000378:metabolism / Q000494:pharmacology; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014764:Viral Proteins; D014779:Virus Replication / Q000187:drug effects; D016335:Zinc Fingers; D054301:gag Gene Products, Human Immunodeficiency Virus
C105419:1,2-dithiane-4,5-diol 1,1-dioxide; D019380:Anti-HIV Agents; D001391:Azo Compounds; D001549:Benzamides; D036022:Capsid Proteins; C100805:DIBA-1; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C494688:GATA1 protein, human; C494689:Gata1 protein, mouse; D015683:Gene Products, gag; C497692:HCFC1 protein, human; C497693:Hcfc1 protein, mouse; D051863:Host Cell Factor C1; D008024:Ligands; C071884:NCP7 protein, Human immunodeficiency virus 1; D009603:Nitroso Compounds; D050811:Octamer Transcription Factor-1; C494165:POU2F1 protein, human; C494166:Pou2f1 protein, mouse; D016329:Sp1 Transcription Factor; D013450:Sulfones; D014157:Transcription Factors; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C004525:azodicarbonamide; C076039:3-nitrosobenzamide; D011065:Poly(ADP-ribose) Polymerases