title: Dynamic modulation of HIV-1 integrase structure and function by cellular lens epithelium-derived growth factor (LEDGF) protein.
abstract: The mandatory integration of the reverse-transcribed HIV-1 genome into host chromatin is catalyzed by the viral protein integrase (IN), and IN activity can be regulated by numerous viral and cellular proteins. Among these, LEDGF has been identified as a cellular cofactor critical for effective HIV-1 integration. The x-ray crystal structure of the catalytic core domain (CCD) of IN in complex with the IN binding domain (IBD) of LEDGF has furthermore revealed essential protein-protein contacts. However, mutagenic studies indicated that interactions between the full-length proteins were more extensive than the contacts observed in the co-crystal structure of the isolated domains. Therefore, we have conducted detailed biochemical characterization of the interactions between full-length IN and LEDGF. Our results reveal a highly dynamic nature of IN subunit-subunit interactions. LEDGF strongly stabilized these interactions and promoted IN tetramerization. Mass spectrometric protein footprinting and molecular modeling experiments uncovered novel intra- and inter-protein-protein contacts in the full-length IN-LEDGF complex that lay outside of the observable IBD-CCD structure. In particular, our studies defined the IN tetramer interface important for enzymatic activities and high affinity LEDGF binding. These findings provide new insight into how LEDGF modulates HIV-1 IN structure and function, and highlight the potential for exploiting the highly dynamic structure of multimeric IN as a novel therapeutic target.
authors.affiliation: Center for Retrovirus Research and Comprehensive Cancer Center, College of Pharmacy, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: McKee
authors.forename: Jacques J
authors.initials: JJ
authors.lastname: Kessl
authors.forename: Nikolozi
authors.initials: N
authors.lastname: Shkriabai
authors.forename: Mohd Jamal
authors.initials: MJ
authors.lastname: Dar
authors.forename: Alan
authors.initials: A
authors.lastname: Engelman
authors.forename: Mamuka
authors.initials: M
authors.lastname: Kvaratskhelia
mesh_terms: D002850:Chromatography, Gel; D019427:HIV Integrase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology*; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D020836:Protein Structure, Quaternary
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; C119380:lens epithelium-derived growth factor; D019427:HIV Integrase
references.citation: J Biol Chem. 1995 Dec 8;270(49):29299-306
references.pmid: 7493962
references.citation: J Biol Chem. 1996 Mar 29;271(13):7712-8
references.pmid: 8631811
references.citation: J Virol. 2003 Apr;77(8):5030-6
references.pmid: 12663813
references.citation: J Biol Chem. 2003 Aug 29;278(35):33528-39
references.pmid: 12796494
references.citation: J Virol. 2004 Apr;78(7):3210-22
references.pmid: 15016842
references.citation: Nat Rev Drug Discov. 2004 Apr;3(4):301-17
references.pmid: 15060526
references.citation: Proc Natl Acad Sci U S A. 2004 May 4;101(18):6894-9
references.pmid: 15118107
references.citation: J Virol. 2004 Sep;78(17):9524-37
references.pmid: 15308744
references.citation: Front Biosci. 2004 Sep 1;9:3187-208
references.pmid: 15353349
references.citation: Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1339-43
references.pmid: 1847518
references.citation: Nucleic Acids Res. 1991 Jul 25;19(14):3821-7
references.pmid: 1861975
references.citation: Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13659-64
references.pmid: 8942990
references.citation: Biochemistry. 1997 Jan 7;36(1):173-80
references.pmid: 8993331
references.citation: Cell. 1997 Feb 21;88(4):483-92
references.pmid: 9038339
references.citation: J Virol. 1997 Jul;71(7):5382-90
references.pmid: 9188609
references.citation: Nat Struct Biol. 1997 Jul;4(7):567-77
references.pmid: 9228950
references.citation: Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9150-4
references.pmid: 9689049
references.citation: Nucleic Acids Res. 1999 May 15;27(10):2202-10
references.pmid: 10219094
references.citation: J Biol Chem. 2004 Nov 19;279(47):48883-92
references.pmid: 15371438
references.citation: J Virol. 2004 Dec;78(23):12735-46
references.pmid: 15542626
references.citation: EMBO J. 2006 Mar 22;25(6):1295-304
references.pmid: 16482214
references.citation: Biochemistry. 2006 Apr 4;45(13):4077-83
references.pmid: 16566581
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 27;103(26):10080-5
references.pmid: 16785440
references.citation: J Mol Biol. 2006 Jul 21;360(4):760-73
references.pmid: 16793062
references.citation: J Biol Chem. 2006 Aug 11;281(32):22707-19
references.pmid: 16774912
references.citation: Science. 2006 Oct 20;314(5798):461-4
references.pmid: 16959972
references.citation: Nucleic Acids Res. 2006;34(18):5157-65
references.pmid: 16998183
references.citation: J Virol. 2006 Dec;80(23):11498-509
references.pmid: 16987986
references.citation: Virology. 2007 Jan 5;357(1):79-90
references.pmid: 16959283
references.citation: J Mol Biol. 2007 Jan 19;365(3):799-811
references.pmid: 17098251
references.citation: J Mol Biol. 2007 Feb 2;365(5):1480-92
references.pmid: 17137594
references.citation: Nucleic Acids Res. 2007;35(1):113-24
references.pmid: 17158150
references.citation: Virology. 2007 Mar 30;360(1):1-5
references.pmid: 17258258
references.citation: Biochemistry. 2007 Mar 13;46(10):2899-908
references.pmid: 17298035
references.citation: J Virol. 2007 Apr;81(8):3969-79
references.pmid: 17267486
references.citation: PLoS Pathog. 2007 Mar;3(3):e47
references.pmid: 17397262
references.citation: Proc Natl Acad Sci U S A. 2007 May 15;104(20):8316-21
references.pmid: 17488811
references.citation: Virology. 2007 Jul 20;364(1):227-36
references.pmid: 17397894
references.citation: Genes Dev. 2007 Jul 15;21(14):1767-78
references.pmid: 17639082
references.citation: Biochemistry. 2007 Oct 9;46(40):11231-9
references.pmid: 17845008
references.citation: PLoS One. 2007;2(12):e1340
references.pmid: 18092005
references.citation: J Biol Chem. 2008 Feb 29;283(9):5632-41
references.pmid: 18093980
references.citation: Antimicrob Agents Chemother. 2008 Apr;52(4):1351-8
references.pmid: 18227187
references.citation: PLoS Pathog. 2008 Mar;4(3):e1000046
references.pmid: 18369482
references.citation: J Biol Chem. 2004 Dec 31;279(53):55570-7
references.pmid: 15475359
references.citation: Biochemistry. 2005 Jan 25;44(3):971-8
references.pmid: 15654753
references.citation: Nucleic Acids Res. 2005;33(3):977-86
references.pmid: 15718297
references.citation: J Cell Sci. 2005 Apr 15;118(Pt 8):1733-43
references.pmid: 15797927
references.citation: Nat Struct Mol Biol. 2005 Jun;12(6):526-32
references.pmid: 15895093
references.citation: J Biol Chem. 2005 Jul 8;280(27):25517-23
references.pmid: 15855167
references.citation: J Biol Chem. 2005 Sep 23;280(38):32775-83
references.pmid: 16006651
references.citation: Proc Natl Acad Sci U S A. 2005 Nov 29;102(48):17308-13
references.pmid: 16260736
references.citation: J Virol. 2006 Feb;80(4):1886-96
references.pmid: 16439544
references.citation: Retrovirology. 2006;3:4
references.pmid: 16409631
references.citation: Nucleic Acids Res. 2006;34(5):1653-65
references.pmid: 16549878
references.citation: FEBS Lett. 2008 Apr 30;582(10):1425-30
references.pmid: 18331842
references.citation: J Virol. 2000 May;74(10):4795-806
references.pmid: 10775618
references.citation: J Virol. 2000 Aug;74(15):7119-26
references.pmid: 10888652
references.citation: Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8233-8
references.pmid: 10890912
references.citation: Biochemistry. 2000 Aug 8;39(31):9275-84
references.pmid: 10924120
references.citation: Acta Crystallogr D Biol Crystallogr. 2001 Apr;57(Pt 4):536-44
references.pmid: 11264582
references.citation: EMBO J. 2001 Jun 15;20(12):3272-81
references.pmid: 11406603
references.citation: EMBO J. 2001 Dec 17;20(24):7333-43
references.pmid: 11743009
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 10;99(25):15988-93
references.pmid: 12461175
references.citation: J Biol Chem. 2003 Jan 3;278(1):372-81
references.pmid: 12407101
references.citation: Bioorg Med Chem Lett. 2003 Mar 24;13(6):1215-9
references.pmid: 12643946
references.citation: J Virol. 1993 Jan;67(1):425-37
references.pmid: 8416376
references.citation: Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3428-32
references.pmid: 8386373
references.citation: Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6125-9
references.pmid: 7687060
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: J Virol. 1994 Aug;68(8):4768-75
references.pmid: 8035478
references.citation: Science. 1994 Dec 23;266(5193):1981-6
references.pmid: 7801124
references.citation: Biochemistry. 1995 Aug 8;34(31):9826-33
references.pmid: 7632683
references.citation: Nat Struct Biol. 1995 Sep;2(9):807-10
references.pmid: 7552753