Human immunodeficiency virus type 1 nucleocapsid protein specifically stimulates Mg2+-dependent DNA integration in vitro.
The integrase (IN) protein of the human immunodeficiency virus mediates integration of the viral DNA into the cellular genome. In vitro, this reaction can be mimicked by using purified recombinant IN and model DNA substrates. IN mediates two reactions: an endonucleolytic cleavage at each 3' end of the proviral DNA (terminal cleavage) and the joining of the linear viral DNA to 5' phosphates in the target DNA (strand transfer). Previous investigators have shown that purified IN requires Mn2+ or Mg2+ to promote strand transfer in vitro, although Mg2+ is the likely metal cofactor in vivo. IN activity in the presence of Mg2+ in vitro requires high IN concentrations and low concentrations of salt. Here, we show that the viral nucleocapsid protein NCp7 allows efficient IN-mediated strand transfer in the presence of Mg2+ at low enzyme concentrations. This potentiating effect appears to be unique to NCp7, as other small DNA-binding proteins, while capable of stimulating integration in the presence of Mn2+, all failed to stimulate strand transfer in the presence of Mg2+.
Carteau
S
S
Laboratoire de Physicochimie et Pharmacologie des Macromolécules Biologiques, CNRS URA 147, Institut Gustave Roussy, Villejuif, France.
Batson
S C
SC
Poljak
L
L
Mouscadet
J F
JF
de Rocquigny
H
H
Darlix
J L
JL
Roques
B P
BP
Käs
E
E
Auclair
C
C
D002213:Capsid / Q000502:physiology*; D036022:Capsid Proteins; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology; D015683:Gene Products, gag / Q000502:physiology*; D015497:HIV-1 / Q000235:genetics*; D019426:Integrases / Q000502:physiology*; D008274:Magnesium / Q000494:pharmacology*; D008345:Manganese / Q000494:pharmacology; D014764:Viral Proteins; D016662:Virus Integration; D054301:gag Gene Products, Human Immunodeficiency Virus
D036022:Capsid Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D015683:Gene Products, gag; C071884:NCP7 protein, Human immunodeficiency virus 1; D014764:Viral Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; D008345:Manganese; D019426:Integrases; D008274:Magnesium