title: Mapping the Vif-A3G interaction using peptide arrays: a basis for anti-HIV lead peptides.
abstract: Human apolipoprotein-B mRNA-editing catalytic polypeptide-like 3G (A3G) is a cytidine deaminase that restricts retroviruses, endogenous retro-elements and DNA viruses. A3G plays a key role in the anti-HIV-1 innate cellular immunity. The HIV-1 Vif protein counteracts A3G mainly by leading A3G towards the proteosomal machinery and by direct inhibition of its enzymatic activity. Both activities involve direct interaction between Vif and A3G. Disrupting the interaction between A3G and Vif may rescue A3G antiviral activity and inhibit HIV-1 propagation. Here, mapping the interaction sites between A3G and Vif by peptide array screening revealed distinct regions in Vif important for A3G binding, including the N-terminal domain (NTD), C-terminal domain (CTD) and residues 83-99. The Vif-binding sites in A3G included 12 different peptides that showed strong binding to either full-length Vif, Vif CTD or both. Sequence similarity was found between Vif-binding peptides from the A3G CTD and NTD. A3G peptides were synthesized and tested for their ability to counteract Vif action. A3G 211-225 inhibited HIV-1 replication in cell culture and impaired Vif dependent A3G degradation. In vivo co-localization of full-length Vif with A3G 211-225 was demonstrated by use of FRET. This peptide has the potential to serve as an anti-HIV-1 lead compound. Our results suggest a complex interaction between Vif and A3G that is mediated by discontinuous binding regions with different affinities.
authors.affiliation: Institute of Chemistry, The Hebrew University of Jerusalem, Safra Campus, Givat Ram, Jerusalem 91904, Israel.
authors.forename: Tali H
authors.initials: TH
authors.lastname: Reingewertz
authors.forename: Elena
authors.initials: E
authors.lastname: Britan-Rosich
authors.forename: Shahar
authors.initials: S
authors.lastname: Rotem-Bamberger
authors.forename: Mathias
authors.initials: M
authors.lastname: Viard
authors.forename: Amy
authors.initials: A
authors.lastname: Jacobs
authors.forename: Abigail
authors.initials: A
authors.lastname: Miller
authors.forename: Ji Youn
authors.initials: JY
authors.lastname: Lee
authors.forename: Jeeseong
authors.initials: J
authors.lastname: Hwang
authors.forename: Robert
authors.initials: R
authors.lastname: Blumenthal
authors.forename: Moshe
authors.initials: M
authors.lastname: Kotler
authors.forename: Assaf
authors.initials: A
authors.lastname: Friedler
mesh_terms: D000071480:APOBEC-3G Deaminase; D019380:Anti-HIV Agents / Q000737:chemistry*; D002478:Cells, Cultured; D003564:Cytidine Deaminase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D006801:Humans; D010449:Peptide Mapping; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D040081:Protein Array Analysis; D054320:vif Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D010455:Peptides; D054320:vif Gene Products, Human Immunodeficiency Virus; C514847:vif protein, Human immunodeficiency virus 1; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase
references.citation: J Virol. 2009 Sep;83(17):8674-82
references.pmid: 19535450
references.citation: Curr HIV Res. 2008 Mar;6(2):91-9
references.pmid: 18336256
references.citation: Nat Rev Immunol. 2004 Nov;4(11):868-77
references.pmid: 15516966
references.citation: Nature. 2008 Mar 6;452(7183):116-9
references.pmid: 18288108
references.citation: J Virol. 2000 Sep;74(18):8252-61
references.pmid: 10954522
references.citation: PLoS Pathog. 2010 Jun 03;6(6):e1000925
references.pmid: 20532212
references.citation: Nat Med. 2003 Nov;9(11):1398-403
references.pmid: 14528301
references.citation: Proc Natl Acad Sci U S A. 2004 Mar 16;101(11):3770-4
references.pmid: 14999100
references.citation: J Virol. 2007 Apr;81(8):3807-15
references.pmid: 17267497
references.citation: J Mol Biol. 2011 Jul 29;410(5):1065-76
references.pmid: 21763507
references.citation: J Mol Biol. 2009 Jun 26;389(5):819-32
references.pmid: 19389408
references.citation: Microbiol Mol Biol Rev. 2009 Jun;73(2):211-32
references.pmid: 19487726
references.citation: Nature. 2002 Aug 8;418(6898):646-50
references.pmid: 12167863
references.citation: J Biol Chem. 2010 May 28;285(22):16546-52
references.pmid: 20363737
references.citation: Mol Cell. 2003 Sep;12(3):591-601
references.pmid: 14527406
references.citation: Proteins. 2007 Nov 1;69(2):270-84
references.pmid: 17598142
references.citation: Bioinformatics. 2007 Nov 1;23(21):2947-8
references.pmid: 17846036
references.citation: Nucleic Acids Res. 2010 Jan;38(2):633-46
references.pmid: 19910370
references.citation: Cell. 2003 Jun 13;113(6):803-9
references.pmid: 12809610
references.citation: Nat Biotechnol. 2008 Oct;26(10):1187-92
references.pmid: 18806783
references.citation: Microbes Infect. 2010 Feb;12(2):166-71
references.pmid: 19944180
references.citation: Nat Struct Mol Biol. 2008 Oct;15(10):1059-66
references.pmid: 18820687
references.citation: Curr Biol. 2003 Nov 11;13(22):2009-13
references.pmid: 14614829
references.citation: Proc Natl Acad Sci U S A. 2004 May 25;101(21):8114-9
references.pmid: 15148397
references.citation: J Biol Chem. 2004 Feb 27;279(9):7792-8
references.pmid: 14672928
references.citation: J Virol. 2000 Oct;74(19):8938-45
references.pmid: 10982337
references.citation: PLoS One. 2010 Jul 12;5(7):e11515
references.pmid: 20635000
references.citation: PLoS One. 2007 Apr 18;2(4):e378
references.pmid: 17440614
references.citation: Curr HIV Res. 2005 Oct;3(4):339-44
references.pmid: 16250885
references.citation: J Virol. 2009 Feb;83(4):1992-2003
references.pmid: 19036809
references.citation: Trends Genet. 2003 Apr;19(4):207-16
references.pmid: 12683974
references.citation: J Virol. 1997 Dec;71(12):9358-65
references.pmid: 9371595
references.citation: Virology. 2006 Jan 20;344(2):263-6
references.pmid: 16303161
references.citation: Nature. 2003 Jul 3;424(6944):94-8
references.pmid: 12808465
references.citation: Retrovirology. 2007 Nov 24;4:81
references.pmid: 18036235
references.citation: PLoS Pathog. 2009 Mar;5(3):e1000330
references.pmid: 19266078
references.citation: J Virol. 2008 Sep;82(17):8656-63
references.pmid: 18562529
references.citation: J Virol. 1997 Jul;71(7):5259-67
references.pmid: 9188594
references.citation: J Virol. 2009 Mar;83(5):2374-81
references.pmid: 19109396
references.citation: FEBS Lett. 1998 Dec 28;441(3):419-26
references.pmid: 9891983
references.citation: J Biol Chem. 2005 Mar 4;280(9):8387-96
references.pmid: 15537645
references.citation: J Biomol NMR. 2001 May;20(1):11-4
references.pmid: 11430750
references.citation: J Mol Biol. 2008 Sep 12;381(4):1000-11
references.pmid: 18619467
references.citation: J Virol. 2007 Aug;81(15):8201-10
references.pmid: 17522216
references.citation: J Biol Chem. 2004 Apr 9;279(15):14481-3
references.pmid: 14966139
references.citation: J Virol. 2007 Dec;81(23):13235-41
references.pmid: 17898068
references.citation: Chem Soc Rev. 2011 May;40(5):2131-45
references.pmid: 21243154
references.citation: J Virol. 2010 Sep;84(17):8561-70
references.pmid: 20592083
references.citation: J Gen Virol. 2002 Sep;83(Pt 9):2225-2230
references.pmid: 12185277
references.citation: J Virol. 2009 Sep;83(17):8544-52
references.pmid: 19535447
references.citation: J Mol Biol. 1999 Mar 19;287(1):93-101
references.pmid: 10074409
references.citation: Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11444-9
references.pmid: 16076960
references.citation: J Clin Virol. 2003 Feb;26(2):143-52
references.pmid: 12600646
references.citation: Genes Dev. 2004 Dec 1;18(23):2861-6
references.pmid: 15574592
references.citation: J Virol. 1992 Apr;66(4):2232-9
references.pmid: 1548759
references.citation: Virology. 2004 Feb 20;319(2):163-75
references.pmid: 15015498
references.citation: Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5652-7
references.pmid: 15054139
references.citation: Trends Pharmacol Sci. 2009 Dec;30(12):638-46
references.pmid: 19837465
references.citation: Proc Natl Acad Sci U S A. 2004 Mar 16;101(11):3927-32
references.pmid: 14978281
references.citation: Protein Pept Lett. 2010 Aug;17(8):988-98
references.pmid: 20450485
references.citation: Antimicrob Agents Chemother. 2005 Sep;49(9):3833-41
references.pmid: 16127060
references.citation: Virology. 2008 Jan 5;370(1):113-21
references.pmid: 17916373
references.citation: Science. 2003 Nov 7;302(5647):1056-60
references.pmid: 14564014
references.citation: Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19539-44
references.pmid: 19887642
references.citation: Virology. 2005 Mar 15;333(2):374-86
references.pmid: 15721369
references.citation: J Virol. 2010 Jun;84(11):5741-50
references.pmid: 20335268
references.citation: Virology. 2008 Jul 20;377(1):49-53
references.pmid: 18499212
references.citation: Mol Immunol. 2007 Jan;44(4):583-90
references.pmid: 16580072
references.citation: Retrovirology. 2009 Jan 07;6:1
references.pmid: 19128510
references.citation: Mol Aspects Med. 2010 Oct;31(5):383-97
references.pmid: 20538015
references.citation: Cell Mol Life Sci. 2009 Oct;66(19):3137-47
references.pmid: 19547914
references.citation: J Biol Chem. 2007 Apr 20;282(16):11667-75
references.pmid: 17272283
references.citation: Protein Eng Des Sel. 2009 May;22(5):281-7
references.pmid: 19218568
references.citation: J Mol Biol. 2007 Oct 26;373(3):541-50
references.pmid: 17869271
references.citation: Virology. 2008 Mar 1;372(1):136-46
references.pmid: 18023836
references.citation: J Virol. 2006 Jun;80(12):5984-91
references.pmid: 16731937
references.citation: Microbes Infect. 2008 Aug-Sep;10(10-11):1218-22
references.pmid: 18617010
references.citation: Nat Biotechnol. 2005 Oct;23(10):1308-14
references.pmid: 16155565
references.citation: Nat Struct Mol Biol. 2012 Oct;19(10):1005-10
references.pmid: 23001005
references.citation: J Biol Chem. 1996 Apr 26;271(17):10121-9
references.pmid: 8626571
references.citation: Anal Biochem. 1989 Nov 1;182(2):319-26
references.pmid: 2610349
references.citation: Nature. 2003 Jul 3;424(6944):99-103
references.pmid: 12808466
references.citation: J Virol. 2003 Nov;77(21):11398-407
references.pmid: 14557625
references.citation: J Biol Chem. 2005 Mar 11;280(10):8765-75
references.pmid: 15611076
references.citation: J Biol Chem. 2006 Jun 23;281(25):17259-17265
references.pmid: 16636053
references.citation: Mol Cell Biol. 2005 Aug;25(16):7270-7
references.pmid: 16055735