The Preserved HTH-Docking Cleft of HIV-1 Integrase Is Functionally Critical.
HIV-1 integrase (IN) catalyzes viral DNA integration into the host genome and facilitates multifunctional steps including virus particle maturation. Competency of IN to form multimeric assemblies is functionally critical, presenting an approach for anti-HIV strategies. Multimerization of IN depends on interactions between the distinct subunit domains and among the flanking protomers. Here, we elucidate an overlooked docking cleft of IN core domain that anchors the N-terminal helix-turn-helix (HTH) motif in a highly preserved and functionally critical configuration. Crystallographic structure of IN core domain in complex with Fab specifically targeting this cleft reveals a steric overlap that would inhibit HTH-docking, C-terminal domain contacts, DNA binding, and subsequent multimerization. While Fab inhibits in vitro IN integration activity, in vivo it abolishes virus particle production by specifically associating with preprocessed IN within Gag-Pol and interfering with early cytosolic Gag/Gag-Pol assemblies. The HTH-docking cleft may offer a fresh hotspot for future anti-HIV intervention strategies.
Department of Biology, Technion - Israel Institute of Technology, Haifa 320003, Israel.
Meytal
M
Galilee
Department of Immunology and Pathology, The Lautenberg Center for General and Tumor Immunology, The Hebrew University Hadassah Medical School, Jerusalem 91120, Israel.
Elena
E
Britan-Rosich
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158, USA.
Sarah L
SL
Griner
Department of Biophysics, Bezmialem Vakif University, Istanbul 34093, Turkey.
Serdar
S
Uysal
Physical Chemistry, Department of Chemistry, Nanosystem Initiative Munich (NIM), Center for Integrated Protein Science Munich (CiPSM), Center for NanoScience (CeNS), Ludwig-Maximilians-Universität München, Munich 81377, Germany.
Viola
V
Baumgärtel
Physical Chemistry, Department of Chemistry, Nanosystem Initiative Munich (NIM), Center for Integrated Protein Science Munich (CiPSM), Center for NanoScience (CeNS), Ludwig-Maximilians-Universität München, Munich 81377, Germany.
Don C
DC
Lamb
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA.
Anthony A
AA
Kossiakoff
Department of Immunology and Pathology, The Lautenberg Center for General and Tumor Immunology, The Hebrew University Hadassah Medical School, Jerusalem 91120, Israel.
Moshe
M
Kotler
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94158, USA.
Robert M
RM
Stroud
Department of Biology, Technion - Israel Institute of Technology, Haifa 320003, Israel.
Ailie
A
Marx
Department of Biology, Technion - Israel Institute of Technology, Haifa 320003, Israel. Electronic address: alian@tx.technion.ac.il.
Akram
A
Alian
D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D019427:HIV Integrase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000737:chemistry / Q000201:enzymology*; D019077:Helix-Turn-Helix Motifs; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D011485:Protein Binding; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D012367:RNA, Viral / Q000378:metabolism
D012367:RNA, Viral; D019427:HIV Integrase
Biophys J. 2013 Aug 20;105(4):848-61
23972837
Curr Top Microbiol Immunol. 2015;389:93-119
25778682
J Cell Biol. 2015 Aug 17;210(4):629-46
26283800
Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
20383002
Nature. 1985 Jan 24-30;313(6000):277-84
2578615
J Biol Chem. 1995 Dec 8;270(49):29299-306
7493962
Nucleic Acids Res. 2008 Dec;36(22):7043-58
18987001
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
19461840
Proc Natl Acad Sci U S A. 1988 Aug;85(16):5879-83
3045807
J Virol. 1992 Apr;66(4):2232-9
1548759
J Biol Chem. 2003 Aug 29;278(35):33528-39
12796494
Retrovirology. 2013 May 30;10:57
23721378
J Biol Chem. 2013 Mar 8;288(10):7373-86
23322775
J Mol Biol. 2007 Nov 2;373(4):924-40
17825836
World J Biol Chem. 2015 Aug 26;6(3):83-94
26322168
PLoS One. 2012;7(10):e47828
23110110
Nature. 2016 Feb 18;530(7590):362-6
26887497
Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9150-4
9689049
Methods. 2009 Apr;47(4):243-8
19010419
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):235-42
21460441
PLoS Pathog. 2009 Jul;5(7):e1000515
19609359
Virology. 2011 Mar 15;411(2):194-205
21216426
Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13659-64
8942990
J Chem Theory Comput. 2015 Sep 8;11(9):4500-11
26575940
J Biol Chem. 2008 Nov 14;283(46):31802-12
18801737
Virology. 2007 Jul 20;364(1):227-36
17397894
J Biol Chem. 2014 Sep 19;289(38):26430-40
25118283
Mol Biol Int. 2012;2012:863405
22928108
J Biol Chem. 1997 Mar 28;272(13):8361-9
9079660
Structure. 2014 Oct 7;22(10):1512-9
25199694
Proc Natl Acad Sci U S A. 2013 May 21;110(21):8690-5
23610442
J Mol Biol. 1993 Feb 20;229(4):969-95
8095303
Proc Natl Acad Sci U S A. 2009 May 19;106(20):8192-7
19416821
PLoS Pathog. 2009 Jan;5(1):e1000259
19132083
Acta Crystallogr D Biol Crystallogr. 2006 Aug;62(Pt 8):859-66
16855301
J Virol. 1997 Oct;71(10):7704-10
9311854
FEBS J. 2009 Jun;276(11):2926-46
19490099
J Biol Chem. 2014 Jul 25;289(30):20477-88
24904063
J Biol Chem. 1996 Mar 29;271(13):7712-8
8631811
Virology. 2013 May 10;439(2):97-104
23473371
J Biol Chem. 2008 Feb 29;283(9):5632-41
18093980
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):355-67
21460454
Mol Ther Methods Clin Dev. 2014 Jan 8;1:null
25383358
PLoS One. 2010 Dec 30;5(12):e16001
21209864
Structure. 2012 Apr 4;20(4):582-92
22483106
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8233-8
10890912
Biochemistry. 2011 Nov 15;50(45):9788-96
21992419
Nature. 2016 Feb 18;530(7590):358-61
26887496
Proc Natl Acad Sci U S A. 2007 May 15;104(20):8316-21
17488811
Nature. 2010 Nov 11;468(7321):326-9
21068843
EMBO J. 2001 Dec 17;20(24):7333-43
11743009
J Virol. 2011 Mar;85(5):2254-65
21159874
J Virol. 2004 Oct;78(19):10803-13
15367647
J Virol. 1995 May;69(5):2729-36
7535863
J Virol. 1996 Jul;70(7):4484-94
8676473
Sci Rep. 2016 Apr 22;6:24957
27102180
Viruses. 2009 Dec;1(3):713-36
21994566
Nature. 2010 Mar 11;464(7286):232-6
20118915