title: Analysis of the anti-apoptotic activity of four vaccinia virus proteins demonstrates that B13 is the most potent inhibitor in isolation and during viral infection.
abstract: Vaccinia virus (VACV) is a large dsDNA virus encoding ~200 proteins, several of which inhibit apoptosis. Here, a comparative study of anti-apoptotic proteins N1, F1, B13 and Golgi anti-apoptotic protein (GAAP) in isolation and during viral infection is presented. VACVs strains engineered to lack each gene separately still blocked apoptosis to some degree because of functional redundancy provided by the other anti-apoptotic proteins. To overcome this redundancy, we inserted each gene separately into a VACV strain (vv811) that lacked all these anti-apoptotic proteins and that induced apoptosis efficiently during infection. Each protein was also expressed in cells using lentivirus vectors. In isolation, each VACV protein showed anti-apoptotic activity in response to specific stimuli, as measured by immunoblotting for cleaved poly(ADP ribose) polymerase-1 and caspase-3 activation. Of the proteins tested, B13 was the most potent inhibitor, blocking both intrinsic and extrinsic stimuli, whilst the activity of the other proteins was largely restricted to inhibition of intrinsic stimuli. In addition, B13 and F1 were effective blockers of apoptosis induced by vv811 infection. Finally, whilst differences in induction of apoptosis were barely detectable during infection with VACV strain Western Reserve compared with derivative viruses lacking individual anti-apoptotic genes, several of these proteins reduced activation of caspase-3 during infection by vv811 strains expressing these proteins. These results illustrated that vv811 was a useful tool to determine the role of VACV proteins during infection and that whilst all of these proteins have some anti-apoptotic activity, B13 was the most potent.
authors.affiliation: Virology Laboratory, Pontchaillou University Hospital, Rennes 35033, France.|Equipe Microbiologie, EA 1254, SFR BIOSIT, Université Européenne de Bretagne, Rennes, France.|Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
authors.forename: David L
authors.initials: DL
authors.lastname: Veyer
authors.affiliation: Department of Virology, Imperial College London, London W2 1PG, UK.|Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
authors.forename: Carlos
authors.initials: C
authors.lastname: Maluquer de Motes
authors.affiliation: Department of Virology, Imperial College London, London W2 1PG, UK.|Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
authors.forename: Rebecca P
authors.initials: RP
authors.lastname: Sumner
authors.affiliation: Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
authors.forename: Louisa
authors.initials: L
authors.lastname: Ludwig
authors.affiliation: Department of Virology, Imperial College London, London W2 1PG, UK.
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Johnson
authors.affiliation: Department of Virology, Imperial College London, London W2 1PG, UK.|Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
authors.forename: Geoffrey L
authors.initials: GL
authors.lastname: Smith
mesh_terms: D017209:Apoptosis; D002460:Cell Line; D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral; D006801:Humans; D012516:Osteosarcoma; D014616:Vaccinia virus / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D014764:Viral Proteins
references.citation: PLoS Pathog. 2008 Feb 8;4(2):e22
references.pmid: 18266467
references.citation: Virology. 1988 May;164(1):121-31
references.pmid: 3284177
references.citation: J Gen Virol. 1999 Apr;80 ( Pt 4):949-959
references.pmid: 10211965
references.citation: Nature. 1995 May 4;375(6526):81-3
references.pmid: 7536901
references.citation: Proc Natl Acad Sci U S A. 2013 May 7;110(19):7808-13
references.pmid: 23603272
references.citation: Virology. 1994 Mar;199(2):491-6
references.pmid: 7510087
references.citation: J Gen Virol. 1989 Sep;70 ( Pt 9):2333-43
references.pmid: 2778436
references.citation: Science. 2010 Feb 12;327(5967):873-876
references.pmid: 20093437
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8318-22
references.pmid: 7667287
references.citation: Nat Rev Mol Cell Biol. 2010 Sep;11(9):621-32
references.pmid: 20683470
references.citation: J Virol. 1996 Sep;70(9):6479-85
references.pmid: 8709286
references.citation: Virology. 1994 Jul;202(1):305-14
references.pmid: 8009842
references.citation: Virology. 1995 Jan 10;206(1):136-47
references.pmid: 7831769
references.citation: Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):14345-50
references.pmid: 14610284
references.citation: Clin Exp Immunol. 1996 Jan;103(1):8-14
references.pmid: 8565292
references.citation: PLoS Pathog. 2007 Feb;3(2):e17
references.pmid: 17319741
references.citation: J Gen Virol. 1997 Mar;78 ( Pt 3):677-85
references.pmid: 9049422
references.citation: PLoS Pathog. 2011 Dec;7(12):e1002430
references.pmid: 22194685
references.citation: Nucleic Acids Res. 1989 Oct 11;17(19):7581-90
references.pmid: 2552411
references.citation: Immunity. 2011 Oct 28;35(4):456-66
references.pmid: 22035838
references.citation: J Virol. 2009 Jul;83(14):7085-98
references.pmid: 19439472
references.citation: Oncogene. 2002 Dec 5;21(55):8379-87
references.pmid: 12466958
references.citation: J Biol Chem. 2012 May 4;287(19):15896-905
references.pmid: 22418439
references.citation: Cell Death Differ. 2006 Oct;13(10):1651-62
references.pmid: 16439990
references.citation: J Gen Virol. 2002 Aug;83(Pt 8):1965-1976
references.pmid: 12124460
references.citation: PLoS Pathog. 2011 Sep;7(9):e1002247
references.pmid: 21931555
references.citation: Semin Immunol. 2001 Feb;13(1):67-72
references.pmid: 11289801
references.citation: PLoS Pathog. 2013 Feb;9(2):e1003183
references.pmid: 23468625
references.citation: Mol Biol Cell. 2009 Aug;20(16):3638-45
references.pmid: 19553469
references.citation: J Biol Chem. 1991 Oct 25;266(30):20103-9
references.pmid: 1657913
references.citation: Protein Sci. 2007 Jan;16(1):118-24
references.pmid: 17123957
references.citation: J Biol Chem. 1995 Feb 17;270(7):3255-60
references.pmid: 7531702
references.citation: J Gen Virol. 2006 Jun;87(Pt 6):1451-1458
references.pmid: 16690909
references.citation: PLoS Pathog. 2014 Apr 17;10(4):e1003989
references.pmid: 24743339
references.citation: J Gen Virol. 2004 Jan;85(Pt 1):105-117
references.pmid: 14718625
references.citation: Virology. 1990 Nov;179(1):247-66, 517-63
references.pmid: 2219722
references.citation: PLoS Pathog. 2008 Aug 15;4(8):e1000128
references.pmid: 18704168
references.citation: J Biol Chem. 2013 May 3;288(18):13057-67
references.pmid: 23508950
references.citation: Cell Death Differ. 2008 Oct;15(10):1564-71
references.pmid: 18551131
references.citation: J Virol. 1997 Mar;71(3):1992-2003
references.pmid: 9032331
references.citation: J Virol. 1995 Dec;69(12):7688-98
references.pmid: 7494278
references.citation: J Gen Virol. 2013 Sep;94(Pt 9):2070-2081
references.pmid: 23761407
references.citation: Proteomics. 2007 Dec;7(23):4228-34
references.pmid: 17979178
references.citation: J Gen Virol. 2013 Nov;94(Pt 11):2367-2392
references.pmid: 23999164
references.citation: J Gen Virol. 2007 Jun;88(Pt 6):1656-1666
references.pmid: 17485524
references.citation: Nat Immunol. 2000 Dec;1(6):489-95
references.pmid: 11101870
references.citation: J Virol. 1984 Mar;49(3):857-64
references.pmid: 6321770
references.citation: J Virol. 1989 Feb;63(2):600-6
references.pmid: 2783466
references.citation: Nat Immunol. 2011 Nov 16;12(12):1143-9
references.pmid: 22089220
references.citation: J Cell Biol. 2013 Aug 19;202(4):699-713
references.pmid: 23940116
references.citation: J Gen Virol. 1991 Jun;72 ( Pt 6):1349-76
references.pmid: 2045793
references.citation: J Biol Chem. 1998 Dec 4;273(49):32608-13
references.pmid: 9829999
references.citation: Cell. 1992 May 15;69(4):597-604
references.pmid: 1339309
references.citation: J Virol. 2001 Dec;75(23):11437-48
references.pmid: 11689625
references.citation: J Virol. 2012 Jan;86(1):203-13
references.pmid: 22013032
references.citation: Nature. 1995 May 4;375(6526):78-81
references.pmid: 7536900
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7829-34
references.pmid: 11427719
references.citation: J Gen Virol. 2012 Oct;93(Pt 10):2098-2108
references.pmid: 22791606
references.citation: Virology. 1991 Jan;180(1):406-10
references.pmid: 1984660
references.citation: Cell. 2009 Jun 12;137(6):1112-23
references.pmid: 19524513
references.citation: J Gen Virol. 2002 Apr;83(Pt 4):855-872
references.pmid: 11907336
references.citation: J Biol Chem. 2004 Aug 27;279(35):36570-8
references.pmid: 15215253
references.citation: J Virol. 2005 Nov;79(22):14031-43
references.pmid: 16254338
references.citation: Virology. 2006 Jan 5;344(1):139-50
references.pmid: 16364745
references.citation: J Virol. 1990 Jun;64(6):3108-11
references.pmid: 2159565