FN Archimer Export Format
PT J
TI A cooperative and specific DNA-binding mode of HIV-1 integrase depends on the nature of the metallic cofactor and involves the zinc-containing N-terminal domain
BT 
AF CARAYON, Kevin
   LEH, Herve
   HENRY, Etienne
   SIMON, Francoise
   MOUSCADET, Jean-Francois
   DEPREZ, Eric
AS 1:1;2:1;3:1;4:1;5:1;6:1;
FF 1:;2:;3:;4:;5:;6:;
C1 Ecole Normale Super, LBPA, CNRS, F-94235 Cachan, France.
C2 Ecole Normale Super, LBPA, CNRS, F-94235 Cachan, France.
IF 7.836
TC 22
UR https://archimer.ifremer.fr/doc/00206/31773/30183.pdf
LA English
DT Article
AB HIV-1 integrase catalyzes the insertion of the viral genome into chromosomal DNA. We characterized the structural determinants of the 3'-processing reaction specificity-the first reaction of the integration process-at the DNA-binding level. We found that the integrase N-terminal domain, containing a pseudo zinc-finger motif, plays a key role, at least indirectly, in the formation of specific integrase-DNA contacts. This motif mediates a cooperative DNA binding of integrase that occurs only with the cognate/viral DNA sequence and the physiologically relevant Mg(2+) cofactor. The DNA-binding was essentially non-cooperative with Mn(2+) or using non-specific/random sequences, regardless of the metallic cofactor. 2,2'-Dithiobisbenzamide-1 induced zinc ejection from integrase by covalently targeting the zinc-finger motif, and significantly decreased the Hill coefficient of the Mg(2+)-mediated integrase-DNA interaction, without affecting the overall affinity. Concomitantly, 2,2'-dithiobisbenzamide-1 severely impaired 3'-processing (IC(50) = 11-15 nM), suggesting that zinc ejection primarily perturbs the nature of the active integrase oligomer. A less specific and weaker catalytic effect of 2,2'-dithiobisbenzamide-1 is mediated by Cys 56 in the catalytic core and, notably, accounts for the weaker inhibition of the non-cooperative Mn(2+)-dependent 3'-processing. Our data show that the cooperative DNA-binding mode is strongly related to the sequence-specific DNA-binding, and depends on the simultaneous presence of the Mg(2+) cofactor and the zinc effector.
PY 2010
PD JUL
SO Nucleic Acids Research
SN 0305-1048
PU Oxford Univ Press
VL 38
IS 11
UT 000279188800026
BP 3692
EP 3708
DI 10.1093/nar/gkq087
ID 31773
ER

EF