Specific recognition of primer tRNA Lys 3 by HIV-1 nucleocapsid protein: involvement of the zinc fingers and the N-terminal basic extension.
Reverse transcription of HIV-1 viral RNA uses human tRNA(Lys)(3) as a primer. We have previously characterised the structural aspects of the tRNA(Lys)(3) interaction by NMR. In HIV-1 virions, the nucleocapsid protein NCp7 contains two zinc fingers flanked by basic residues. Using NMR, the respective role of the N-terminal residues and the zinc fingers in the interaction with tRNA(Lys)(3)/NCp7 was investigated by studying the tRNA(Lys)(3) and tRNA(Lys)(3)/NCp7 complexes. The N-terminal basic residues do not change the footprint of NC on tRNA(Lys)(3)/Cys(23)NCp7 but strengthen the binding whereas the mutation of the zinc-binding histidine at position 23 that was shown to result in non-infectious particles prevents the interaction with the first bases of the acceptor stem.
Tisné
Carine
C
Laboratoire de Cristallographie et RMN Biologiques, UMR 8015 CNRS, Faculté de Pharmacie, 4, avenue de l'Observatoire, 75006 Paris, France. tisne@pharmacie.univ-paris5.fr
Roques
Bernard P
BP
Dardel
Frédéric
F
D000818:Animals; D036022:Capsid Proteins / Q000737:chemistry / Q000502:physiology*; D015683:Gene Products, gag / Q000737:chemistry / Q000502:physiology*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D008279:Magnetic Resonance Imaging; D009154:Mutation; D011485:Protein Binding; D012357:RNA, Transfer, Lys / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins; D014779:Virus Replication / Q000502:physiology; D016335:Zinc Fingers / Q000502:physiology; D054301:gag Gene Products, Human Immunodeficiency Virus
D036022:Capsid Proteins; D015683:Gene Products, gag; C071884:NCP7 protein, Human immunodeficiency virus 1; D012357:RNA, Transfer, Lys; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus