title: Identification of novel key amino acids at the interface of the transmembrane domains of human BST-2 and HIV-1 Vpu.
abstract: BACKGROUND
  BST-2 (bone marrow stromal cell antigen 2) is an interferon-inducible protein that inhibits virus release by tethering viral particles to the cell surface. This antiviral activity of BST-2 is antagonized by HIV-1 accessory protein Vpu. Vpu physically interacts with BST-2 through their mutual transmembrane (TM) domains. In this study, we utilized the BRET assay and molecular dynamics (MD) simulation method to further characterize the interaction of BST-2 and Vpu.
  
  
  RESULTS
  Amino acids I34, L37, P40 and L41 in the TM domain of BST-2, and L11, A18 and W22 in the TM domain of Vpu were identified to be critical for the interaction between BST-2 and Vpu. The residues P40 in the TM domain of BST-2 and L11 in the TM domain of Vpu were shown, for the first time, to be important for their interaction. Furthermore, triple-amino-acid substitutions, 14-16 (AII to VAA) and 26-28 (IIE to AAA) in Vpu TM, not the single-residue mutation, profoundly disrupted BST-2/Vpu interaction. The results of MD simulation revealed significant conformational changes of the BST-2/Vpu complex as a result of mutating P40 of BST-2 and L11, 14-16 (AII to VAA) and 26-28 (IIE to AAA) of Vpu. In addition, disrupting the interaction between BST-2 and Vpu rendered BST-2 resistant to Vpu antagonization.
  
  
  CONCLUSIONS
  Through use of the BRET assay, we identified novel key residues P40 in the TM domain of BST-2 and L11 in the TM domain of Vpu that are important for their interaction. These results add new insights into the molecular mechanism behind BST-2 antagonization by HIV-1 Vpu.
authors.affiliation: MOH Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Beijing 100730, P. R. China.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Pang
authors.forename: Siqi
authors.initials: S
authors.lastname: Hu
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.forename: Fengwen
authors.initials: F
authors.lastname: Xu
authors.forename: Shan
authors.initials: S
authors.lastname: Mei
authors.forename: Jinming
authors.initials: J
authors.lastname: Zhou
authors.forename: Shan
authors.initials: S
authors.lastname: Cen
authors.forename: Qi
authors.initials: Q
authors.lastname: Jin
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
mesh_terms: D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism*; D060807:Bioluminescence Resonance Energy Transfer Techniques; D002460:Cell Line; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D054298:Human Immunodeficiency Virus Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics* / Q000378:metabolism*; D000074010:Bone Marrow Stromal Antigen 2; D000086082:Viroporin Proteins
chemical_list: D015703:Antigens, CD; D058851:GPI-Linked Proteins; D054298:Human Immunodeficiency Virus Proteins; D054334:Viral Regulatory and Accessory Proteins; C094572:BST2 protein, human; D000074010:Bone Marrow Stromal Antigen 2; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins
references.citation: PLoS Pathog. 2009 Feb;5(2):e1000300
references.pmid: 19214216
references.citation: J Virol. 2009 Aug;83(15):7536-46
references.pmid: 19474106
references.citation: PLoS One. 2011;6(6):e20890
references.pmid: 21674066
references.citation: Retrovirology. 2012 Jan 27;9:10
references.pmid: 22284121
references.citation: Nature. 2008 Jan 24;451(7177):425-30
references.pmid: 18200009
references.citation: Front Microbiol. 2011 Dec 12;2:250
references.pmid: 22180752
references.citation: Virol J. 2011 May 02;8:198
references.pmid: 21529378
references.citation: PLoS Pathog. 2009 May;5(5):e1000450
references.pmid: 19478868
references.citation: Methods Mol Biol. 2004;259:323-33
references.pmid: 15250502
references.citation: PLoS Pathog. 2010 Feb 05;6(2):e1000749
references.pmid: 20140192
references.citation: Traffic. 2003 Oct;4(10):694-709
references.pmid: 12956872
references.citation: Cell Host Microbe. 2008 Apr 17;3(4):245-52
references.pmid: 18342597
references.citation: Proc Natl Acad Sci U S A. 1999 Jan 5;96(1):151-6
references.pmid: 9874787
references.citation: J Virol. 2010 Dec;84(24):12958-70
references.pmid: 20926557
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6798-802
references.pmid: 15084749
references.citation: J Virol. 2010 Dec;84(23):12336-43
references.pmid: 20861258
references.citation: Antiviral Res. 2011 Sep;91(3):321-9
references.pmid: 21777622
references.citation: PLoS Pathog. 2009 Sep;5(9):e1000574
references.pmid: 19730691
references.citation: J Biol Chem. 2012 Jan 2;287(1):58-67
references.pmid: 22072710
references.citation: AIDS Res Hum Retroviruses. 2009 Dec;25(12):1197-210
references.pmid: 19929170
references.citation: J Virol. 2010 May;84(9):4415-25
references.pmid: 20181686
references.citation: Nat Methods. 2006 Mar;3(3):165-74
references.pmid: 16489332
references.citation: Biochem Biophys Res Commun. 2012 Nov 9;428(1):17-23
references.pmid: 23047007
references.citation: J Virol. 2011 Jan;85(2):932-45
references.pmid: 21068238
references.citation: PLoS One. 2011;6(11):e27660
references.pmid: 22110710
references.citation: J Virol. 2009 Aug;83(16):7931-47
references.pmid: 19515779
references.citation: Biochemistry. 2012 Feb 14;51(6):1288-96
references.pmid: 22264038
references.citation: J Virol. 2009 Feb;83(4):1837-44
references.pmid: 19036818
references.citation: PLoS Pathog. 2010 Mar 05;6(3):e1000701
references.pmid: 20221443
references.citation: PLoS One. 2010 Feb 22;5(2):e9344
references.pmid: 20179761
references.citation: Virology. 2011 Mar 1;411(1):65-77
references.pmid: 21237475
references.citation: Cell. 2009 Oct 30;139(3):499-511
references.pmid: 19879838
references.citation: Nat Protoc. 2006;1(1):337-45
references.pmid: 17406254
references.citation: Curr HIV Res. 2012 Jun;10(4):292-7
references.pmid: 22524177
references.citation: PLoS Pathog. 2013;9(4):e1003299
references.pmid: 23633949
references.citation: PLoS Pathog. 2009 May;5(5):e1000443
references.pmid: 19461879
references.citation: Science. 1988 Sep 2;241(4870):1221-3
references.pmid: 3261888