title: Epstein-Barr Virus SM protein utilizes cellular splicing factor SRp20 to mediate alternative splicing.
abstract: Epstein-Barr virus (EBV) SM protein is an essential nuclear protein produced during the lytic cycle of EBV replication. SM is an RNA-binding protein with multiple mechanisms of action. SM enhances the expression of EBV genes by stabilizing mRNA and facilitating nuclear export. SM also influences splicing of both EBV and cellular pre-mRNAs. SM modulates splice site selection of the host cell STAT1 pre-mRNA, directing utilization of a novel 5' splice site that is used only in the presence of SM. SM activates splicing in the manner of SR proteins but does not contain the canonical RS domains typical of cellular splicing factors. Affinity purification and mass spectrometry of SM complexes from SM-transfected cells led to the identification of the cellular SR splicing factor SRp20 as an SM-interacting protein. The regions of SM and SRp20 required for interaction were mapped by in vitro and in vivo assays. The SRp20 interaction was shown to be important for the effects of SM on alternative splicing by the use of STAT1 splicing assays. Overexpression of SRp20 enhanced SM-mediated alternative splicing and knockdown of SRp20 inhibited the SM effect on splicing. These data suggest a model whereby SM, a viral protein, recruits and co-opts the function of cellular SRp20 in alternative splicing.
authors.affiliation: Department of Medicine, University of Florida, Gainesville, FL 32610, USA.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Verma
authors.forename: Swarna
authors.initials: S
authors.lastname: Bais
authors.forename: Melusine
authors.initials: M
authors.lastname: Gaillard
authors.forename: Sankar
authors.initials: S
authors.lastname: Swaminathan
mesh_terms: D017398:Alternative Splicing; D002460:Cell Line; D020031:Epstein-Barr Virus Infections / Q000378:metabolism* / Q000821:virology; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D016601:RNA-Binding Proteins; D000068103:Serine-Arginine Splicing Factors; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*
chemical_list: C068382:BRLF1 protein, Human herpesvirus 4; D017874:Immediate-Early Proteins; D016601:RNA-Binding Proteins; C491388:SRSF3 protein, human; D015534:Trans-Activators; D000068103:Serine-Arginine Splicing Factors
references.citation: J Biol Chem. 1997 Nov 7;272(45):28582-9
references.pmid: 9353323
references.citation: J Virol. 2002 Oct;76(19):9635-44
references.pmid: 12208942
references.citation: J Virol. 1999 May;73(5):4090-100
references.pmid: 10196305
references.citation: J Virol. 2003 Mar;77(6):3690-701
references.pmid: 12610144
references.citation: J Virol. 1998 Nov;72(11):8485-92
references.pmid: 9765385
references.citation: J Virol. 2000 Jul;74(13):6068-76
references.pmid: 10846090
references.citation: J Virol. 2009 Nov;83(22):11635-44
references.pmid: 19726500
references.citation: J Virol. 2009 Aug;83(16):8041-50
references.pmid: 19515786
references.citation: Science. 2003 Dec 19;302(5653):2141-4
references.pmid: 14684825
references.citation: J Gen Virol. 1977 Jul;36(1):59-74
references.pmid: 886304
references.citation: J Proteome Res. 2004 May-Jun;3(3):463-8
references.pmid: 15253427
references.citation: PLoS Pathog. 2010 Mar 05;6(3):e1000799
references.pmid: 20221435
references.citation: Nat Rev Mol Cell Biol. 2009 Nov;10(11):741-54
references.pmid: 19773805
references.citation: Mol Cell. 2009 Apr 24;34(2):179-90
references.pmid: 19394295
references.citation: J Virol. 2001 Jul;75(13):6033-41
references.pmid: 11390605
references.citation: Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8852-7
references.pmid: 9671768
references.citation: J Virol. 1982 Dec;44(3):834-44
references.pmid: 6294333
references.citation: Nat Struct Mol Biol. 2007 Jul;14(7):597-603
references.pmid: 17603499
references.citation: J Med Virol. 2003 Mar;69(3):306-12
references.pmid: 12526039
references.citation: Blood. 2004 Oct 15;104(8):2475-83
references.pmid: 15217838
references.citation: Proc Natl Acad Sci U S A. 2008 Apr 1;105(13):5154-9
references.pmid: 18364396
references.citation: Science. 1997 Sep 12;277(5332):1630-5
references.pmid: 9287210
references.citation: J Virol. 2009 Dec;83(24):12759-68
references.pmid: 19793817
references.citation: Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9666-70
references.pmid: 15210956
references.citation: J Virol. 1998 Dec;72(12):9526-34
references.pmid: 9811685
references.citation: EMBO J. 2006 Nov 1;25(21):5126-37
references.pmid: 17036044
references.citation: EMBO J. 2003 Apr 1;22(7):1608-19
references.pmid: 12660167
references.citation: J Virol. 2004 Sep;78(17):9412-22
references.pmid: 15308735
references.citation: J Biol Chem. 2003 Jan 3;278(1):335-42
references.pmid: 12403791
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 16;101(46):16286-91
references.pmid: 15534216
references.citation: J Biol Chem. 2003 Sep 26;278(39):37790-8
references.pmid: 12857728
references.citation: Genes Dev. 2006 Jul 1;20(13):1755-65
references.pmid: 16766678
references.citation: Mol Cell Biol. 1998 Apr;18(4):2205-17
references.pmid: 9528792
references.citation: Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7673-8
references.pmid: 8755534
references.citation: J Virol. 1999 Aug;73(8):6872-81
references.pmid: 10400785
references.citation: Virology. 2010 Jun 5;401(2):155-64
references.pmid: 20227104
references.citation: J Virol. 2007 Apr;81(8):4058-69
references.pmid: 17287267
references.citation: RNA. 2000 Sep;6(9):1197-211
references.pmid: 10999598
references.citation: Mol Cell Biol. 1996 Jan;16(1):288-93
references.pmid: 8524306
references.citation: Mol Cell Biol. 1998 Sep;18(9):4977-85
references.pmid: 9710581
references.citation: J Virol. 2008 Jul;82(14):7180-8
references.pmid: 18463151