title: HIV-1 integrase crosslinked oligomers are active in vitro.
abstract: 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.
authors.affiliation: CNRS UMR 5097, Université Victor Segalen Bordeaux 2, IFR 66 Pathologies Infectieuses et Cancers 146 rue Léo Saignat, 33076 Bordeaux cedex, France.
authors.forename: Aurélie
authors.initials: A
authors.lastname: Faure
authors.forename: Christina
authors.initials: C
authors.lastname: Calmels
authors.forename: Cécile
authors.initials: C
authors.lastname: Desjobert
authors.forename: Michel
authors.initials: M
authors.lastname: Castroviejo
authors.forename: Anne
authors.initials: A
authors.lastname: Caumont-Sarcos
authors.forename: Laura
authors.initials: L
authors.lastname: Tarrago-Litvak
authors.forename: Simon
authors.initials: S
authors.lastname: Litvak
authors.forename: Vincent
authors.initials: V
authors.lastname: Parissi
mesh_terms: 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
chemical_list: D003432:Cross-Linking Reagents; D011994:Recombinant Proteins; D004247:DNA; D019427:HIV Integrase
references.citation: EMBO J. 2001 Jul 2;20(13):3565-76
references.pmid: 11432843
references.citation: J Virol. 1995 Dec;69(12):7483-8
references.pmid: 7494254
references.citation: J Virol. 2001 Dec;75(23):11344-53
references.pmid: 11689615
references.citation: EMBO J. 2001 Dec 17;20(24):7333-43
references.pmid: 11743009
references.citation: J Virol. 2002 Apr;76(7):3105-13
references.pmid: 11884535
references.citation: J Biol Chem. 2002 Oct 11;277(41):38045-52
references.pmid: 12147698
references.citation: J Biol Chem. 2003 Jan 3;278(1):372-81
references.pmid: 12407101
references.citation: J Biol Chem. 2003 Jan 10;278(2):1323-7
references.pmid: 12446721
references.citation: Cell. 1990 Oct 5;63(1):87-95
references.pmid: 2170022
references.citation: Microbiol Mol Biol Rev. 1999 Dec;63(4):836-43, table of contents
references.pmid: 10585967
references.citation: Science. 2000 Jan 28;287(5453):646-50
references.pmid: 10649997
references.citation: Science. 2000 Jul 7;289(5476):77-85
references.pmid: 10884228
references.citation: Biochemistry. 2000 Aug 8;39(31):9275-84
references.pmid: 10924120
references.citation: Biochemistry. 2000 Aug 8;39(31):9285-94
references.pmid: 10924121
references.citation: Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11244-9
references.pmid: 11016953
references.citation: J Biol Chem. 2001 Jun 29;276(26):23213-6
references.pmid: 11346660
references.citation: Biopolymers. 2003 Jan;68(1):110-20
references.pmid: 12579583
references.citation: Eur J Biochem. 2003 Nov;270(22):4426-38
references.pmid: 14622271
references.citation: Nucleic Acids Res. 2004;32(4):1527-38
references.pmid: 14999095
references.citation: J Mol Biol. 1983 Jun 5;166(4):557-80
references.pmid: 6345791
references.citation: Science. 1985 Feb 1;227(4686):484-92
references.pmid: 2578227
references.citation: Methods Enzymol. 1985;114:147-56
references.pmid: 4079763
references.citation: Cell. 1990 Aug 24;62(4):829-37
references.pmid: 2167180
references.citation: EMBO J. 1991 Oct;10(10):3051-9
references.pmid: 1655409
references.citation: J Virol. 1992 Aug;66(8):5092-5
references.pmid: 1629963
references.citation: Cell. 1992 Jul 24;70(2):303-11
references.pmid: 1322248
references.citation: EMBO J. 1993 Aug;12(8):3261-7
references.pmid: 8344263
references.citation: EMBO J. 1993 Aug;12(8):3269-75
references.pmid: 8344264
references.citation: Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7316-20
references.pmid: 8041787
references.citation: J Virol. 1995 Oct;69(10):6090-7
references.pmid: 7666512
references.citation: J Virol. 1996 Jun;70(6):3571-80
references.pmid: 8648691
references.citation: Curr Genet. 1996 May;29(6):503-10
references.pmid: 8662188
references.citation: Biochemistry. 1997 Jan 7;36(1):173-80
references.pmid: 8993331
references.citation: Eur J Biochem. 1998 Jan 15;251(1-2):487-95
references.pmid: 9492322
references.citation: Biochemistry. 1998 May 12;37(19):6667-78
references.pmid: 9578550
references.citation: Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10535-40
references.pmid: 9724738
references.citation: J Biol Chem. 1998 Dec 25;273(52):35078-87
references.pmid: 9857042
references.citation: J Virol. 1999 Apr;73(4):3176-83
references.pmid: 10074170
references.citation: J Virol. 1999 Aug;73(8):6670-9
references.pmid: 10400764
references.citation: FEBS Lett. 1999 Jul 16;455(1-2):154-8
references.pmid: 10428491
references.citation: Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10090-5
references.pmid: 11504911