HIV-1 integrase crosslinked oligomers are active in vitro.
The oligomeric state of active human immunodeficiency virus type 1 (HIV-1) integrase (IN) has not been clearly elucidated. We analyzed the activity of the different purified oligomeric forms of recombinant IN obtained after stabilization by platinum crosslinking. The crosslinked tetramer isolated by gel chromatography was able to catalyze the full-site integration of the two viral LTR ends into a target DNA in vitro, whereas the isolated dimeric form of the enzyme was involved in the processing and integration of only one viral end. Accurate concerted integration by IN tetramers was confirmed by cloning and sequencing. Kinetic studies of DNA-integrase complexes led us to propose a model explaining the formation of an active complex. Our data suggest that the tetrameric IN bound to the viral DNA ends is the minimal complex involved in the concerted integration of both LTRs and should be the oligomeric form targeted by future inhibitors.
Faure
Aurélie
A
CNRS UMR 5097, Université Victor Segalen Bordeaux 2, IFR 66 Pathologies Infectieuses et Cancers 146 rue Léo Saignat, 33076 Bordeaux cedex, France.
Calmels
Christina
C
Desjobert
Cécile
C
Castroviejo
Michel
M
Caumont-Sarcos
Anne
A
Tarrago-Litvak
Laura
L
Litvak
Simon
S
Parissi
Vincent
V
D003432:Cross-Linking Reagents; D004247:DNA / Q000378:metabolism; D019427:HIV Integrase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000201:enzymology*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015003:Yeasts / Q000235:genetics
D003432:Cross-Linking Reagents; D011994:Recombinant Proteins; D004247:DNA; D019427:HIV Integrase
EMBO J. 2001 Jul 2;20(13):3565-76
11432843
J Virol. 1995 Dec;69(12):7483-8
7494254
J Virol. 2001 Dec;75(23):11344-53
11689615
EMBO J. 2001 Dec 17;20(24):7333-43
11743009
J Virol. 2002 Apr;76(7):3105-13
11884535
J Biol Chem. 2002 Oct 11;277(41):38045-52
12147698
J Biol Chem. 2003 Jan 3;278(1):372-81
12407101
J Biol Chem. 2003 Jan 10;278(2):1323-7
12446721
Cell. 1990 Oct 5;63(1):87-95
2170022
Microbiol Mol Biol Rev. 1999 Dec;63(4):836-43, table of contents
10585967
Science. 2000 Jan 28;287(5453):646-50
10649997
Science. 2000 Jul 7;289(5476):77-85
10884228
Biochemistry. 2000 Aug 8;39(31):9275-84
10924120
Biochemistry. 2000 Aug 8;39(31):9285-94
10924121
Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11244-9
11016953
J Biol Chem. 2001 Jun 29;276(26):23213-6
11346660
Biopolymers. 2003 Jan;68(1):110-20
12579583
Eur J Biochem. 2003 Nov;270(22):4426-38
14622271
Nucleic Acids Res. 2004;32(4):1527-38
14999095
J Mol Biol. 1983 Jun 5;166(4):557-80
6345791
Science. 1985 Feb 1;227(4686):484-92
2578227
Methods Enzymol. 1985;114:147-56
4079763
Cell. 1990 Aug 24;62(4):829-37
2167180
EMBO J. 1991 Oct;10(10):3051-9
1655409
J Virol. 1992 Aug;66(8):5092-5
1629963
Cell. 1992 Jul 24;70(2):303-11
1322248
EMBO J. 1993 Aug;12(8):3261-7
8344263
EMBO J. 1993 Aug;12(8):3269-75
8344264
Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7316-20
8041787
J Virol. 1995 Oct;69(10):6090-7
7666512
J Virol. 1996 Jun;70(6):3571-80
8648691
Curr Genet. 1996 May;29(6):503-10
8662188
Biochemistry. 1997 Jan 7;36(1):173-80
8993331
Eur J Biochem. 1998 Jan 15;251(1-2):487-95
9492322
Biochemistry. 1998 May 12;37(19):6667-78
9578550
Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10535-40
9724738
J Biol Chem. 1998 Dec 25;273(52):35078-87
9857042
J Virol. 1999 Apr;73(4):3176-83
10074170
J Virol. 1999 Aug;73(8):6670-9
10400764
FEBS Lett. 1999 Jul 16;455(1-2):154-8
10428491
Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10090-5
11504911