S. Carteau et al., HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 NUCLEOCAPSID PROTEIN SPECIFICALLYSTIMULATES MG2-DEPENDENT DNA INTEGRATION IN-VITRO(), Journal of virology, 71(8), 1997, pp. 6225-6229
The integrase (IN) protein of the human immunodeficiency virus mediate
s integration of the viral DNA into the cellular genome. In vitro, thi
s 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 joinin
g of the linear viral DNA to 5' phosphates in the target DNA (strand t
ransfer). 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 efficie
nt IN-mediated strand transfer in the presence of Mg2+ at low enzyme c
oncentrations. This potentiating effect appears to be unique to NCp7,
as other smalt DNA-binding proteins, while capable of stimulating inte
gration in the presence of Mn2+, all failed to stimulate strand transf
er in the presence of Mg2+.