Identification of interaction between HIV-1 glycoprotein 41 and integrase.
Human immunodeficiency virus-1 (HIV-1) encodes 15 viral proteins. Protein-protein interactions play a large role in the function of these proteins. In this study, we attempted to identify novel interactions between the HIV-1 proteins to better understand the role played by viral protein-protein interactions in the life cycle of HIV-1. Genes encoding the 15 viral proteins from the HIV-1 strain AD8 were inserted into the plasmids of a yeast two-hybrid system. By screening 120 pairs of proteins, interactions between seven pairs were found. This led to the discovery of an interaction between the HIV-1 proteins integrase (IN) and glycoprotein 41 (gp41), which was confirmed by both co-immunoprecipitation (Co-IP) assays and fluorescence resonance energy transfer (FRET) imaging in live cells. In addition, it was found that the amino acids at positions 76-100 of gp41 are required for it to bind to IN. Deletion of this region from gp41 prevented its interaction with IN and reduced the production of HIV-1 in 293T cells. This study provides new information on HIV-1 protein-protein interactions which improves the understanding of the biological functions of gp41 and IN during the virus life cycle.
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Xiaowei
X
Zhang
College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
Fei
F
Zhang
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Xiaohe
X
Ma
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Xing
X
Zhao
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Wei
W
Li
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China.
Zhiping
Z
Zhang
College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
Jibin
J
Zhang
National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. czq@wh.iov.cn.
Xian-En
XE
Zhang
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China. zhangxe@ibp.ac.cn.
Zongqiang
Z
Cui
D020816:Amino Acid Motifs; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015658:HIV Infections / Q000821:virology; D019427:HIV Integrase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D020798:Two-Hybrid System Techniques
D015700:HIV Envelope Protein gp41; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1
glycoprotein 41 (gp41); human immunodeficiency virus-1 (HIV-1); integrase (IN); protein-protein interactions; yeast two-hybrid assay
Proc Natl Acad Sci U S A. 2014 Mar 4;111(9):3425-30
24550514
Nature. 1994 Jan 20;367(6460):295-9
8121496
Lancet. 2015 Jul 11;386(9989):171-218
26117719
Cell. 1997 Apr 18;89(2):263-73
9108481
J Mol Biol. 2007 Jun 22;369(5):1230-43
17490682
Biochim Biophys Acta. 2001 May 31;1536(2-3):116-22
11406346
J Biol Chem. 1997 Dec 5;272(49):30753-9
9388214
J Virol. 2015 Dec;89(23):12058-69
26401032
Cell. 1998 May 29;93(5):685-92
9630214
J Virol. 2000 Apr;74(8):3548-54
10729129
Nat Struct Mol Biol. 2004 Jul;11(7):672-5
15208690
Annu Rev Genet. 1999;33:133-70
10690406
Nat Med. 2007 Oct;13(10):1241-7
17906637
Nature. 1996 Nov 14;384(6605):179-83
8906795
J Microsc. 2003 Jan;209(Pt 1):56-70
12535185
Nat Rev Microbiol. 2013 Dec;11(12 ):877-83
24162027
Chem Soc Rev. 2011 Mar;40(3):1337-46
21152581
J Virol. 2004 May;78(10):5056-67
15113887
Trends Microbiol. 2004 Apr;12(4):170-7
15051067
EMBO J. 1998 Aug 17;17(16):4572-84
9707417
J Virol. 2001 Nov;75(21):10537-42
11581428
J Virol. 1997 Dec;71(12):9722-31
9371638
Nat Med. 2001 Aug;7(8):920-6
11479624
Biochem J. 2002 Feb 1;361(Pt 3):557-66
11802785
J Biol Chem. 2007 May 25;282(21):15743-53
17403681
Biotechniques. 2004 May;36(5):872-6
15152608
Microbiol Rev. 1993 Mar;57(1):183-289
8464405
Science. 1998 Jun 19;280(5371):1884-8
9632381
J Virol. 1999 Jan;73(1):592-600
9847364
Cell. 2008 May 16;133(4):742, 742.e1
18485880
Nature. 1988 Jan 21;331(6153):280-3
2447506
Nat Rev Microbiol. 2007 Apr;5(4):253-63
17325726
Nat Rev Microbiol. 2014 Nov;12(11):750-64
25402363
J Biol Chem. 2009 Mar 20;284(12):7931-9
19150986
Blood. 2005 Apr 1;105(7):2802-11
15591114
Cell. 2008 Oct 3;135(1):49-60
18854154
Protein Eng Des Sel. 2009 Dec;22(12):753-63
19875511
Nature. 1997 Aug 28;388(6645):882-7
9278050
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8233-8
10890912
J Virol. 1994 Mar;68(3):1633-42
8107224
Virology. 2012 Jan 20;422(2):265-77
22104209
Nat Med. 2005 Dec;11(12):1287-9
16311605
J Virol. 1996 Oct;70(10):6820-5
8794322
Cell Host Microbe. 2008 Nov 13;4(5):495-504
18976975
J Med Chem. 2010 Jul 22;53(14):5356-60
20586421
Methods Enzymol. 1983;101:181-91
6310321
Annu Rev Biochem. 1998;67:1-25
9759480