Viral RNA annealing activities of human immunodeficiency virus type 1 nucleocapsid protein require only peptide domains outside the zinc fingers.
The nucleocapsid (NC) of human immunodeficiency virus type 1 consists of a large number of NC protein molecules, probably wrapping the dimeric RNA genome within the virion inner core. NC protein is a gag-encoded product that contains two zinc fingers flanked by basic residues. In human immunodeficiency virus type 1 virions, NCp15 is ultimately processed into NCp7 and p6 proteins. During virion assembly the retroviral NC protein is necessary for core formation and genomic RNA encapsidation, which are essential for virus infectivity. In vitro NCp15 activates viral RNA dimerization, a process most probably linked in vivo to genomic RNA packaging, and replication primer tRNA(Lys,3) annealing to the initiation site of reverse transcription. To characterize the domains of human immunodeficiency virus type 1 NC protein necessary for its various functions, the 72-amino acid NCp7 and several derived peptides were synthesized in a pure form. We show here that synthetic NCp7 with or without the two zinc fingers has the RNA annealing activities of NCp15. Further deletions of the N-terminal 12 and C-terminal 8 amino acids, leading to a 27-residue peptide lacking the finger domains, have little or no effect on NC protein activity in vitro. However deletion of short sequences containing basic residues flanking the first finger leads to a complete loss of NC protein activity. It is proposed that the basic residues and the zinc fingers cooperate to select and package the genomic RNA in vivo. Inhibition of the viral RNA binding and annealing activities associated with the basic residues flanking the first zinc finger of NC protein could therefore be used as a model for the design of antiviral agents.
De Rocquigny
H
H
Département de Chimie Organique, U266 Institut National de la Santé et de la Recherche Médicale Unité Associeé 498, Université René Descartes, Paris, France.
Gabus
C
C
Vincent
A
A
Fournié-Zaluski
M C
MC
Roques
B
B
Darlix
J L
JL
D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D015683:Gene Products, gag / Q000378:metabolism*; D015497:HIV-1 / Q000648:ultrastructure*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism; D011485:Protein Binding; D012357:RNA, Transfer, Lys / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000648:ultrastructure; D014758:Viral Core Proteins / Q000737:chemistry / Q000378:metabolism*; D016335:Zinc Fingers
D015683:Gene Products, gag; D010455:Peptides; D012357:RNA, Transfer, Lys; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014758:Viral Core Proteins