title: Characterization of nuclear localization signals (NLSs) and function of NLSs and phosphorylation of serine residues in subcellular and subnuclear localization of transformer-2β (Tra2β).
abstract: The serine/arginine-rich (SR) proteins are one type of major actors in regulation of pre-mRNA splicing. Their functions are closely related to the intracellular spatial organization. The RS domain and phosphorylation status of SR proteins are two critical factors in determining the subcellular distribution. Mammalian Transformer-2β (Tra2β) protein, a member of SR proteins, is known to play multiple important roles in development and diseases. In the present study, we characterized the subcellular and subnuclear localization of Tra2β protein and its related mechanisms. The results demonstrated that in the brain the nuclear and cytoplasmic localization of Tra2β were correlated with its phosphorylation status. Using deletional mutation analysis, we showed that the nuclear localization of Tra2β was determined by multiple nuclear localization signals (NLSs) in the RS domains. The point-mutation analysis disclosed that phosphorylation of serine residues in the NLSs inhibited the function of NLS in directing Tra2β to the nucleus. In addition, we identified at least two nuclear speckle localization signals within the RS1 domain, but not in the RS2 domain. The nuclear speckle localization signals determined the localization of RS1 domain-contained proteins to the nuclear speckle. The function of the signals did not depend on the presence of serine residues. The results provide new insight into the mechanisms by which the subcellular and subnuclear localization of Tra2β proteins are regulated.
authors: Laboratory of Genomic Physiology, State Key Laboratory of Medical Neurobiology, Institutes of Brain Science, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai, 200032 China.
authors: Shu-Jing
authors: SJ
authors: Li
authors: Yao
authors: Y
authors: Qi
authors: Jing-Jing
authors: JJ
authors: Zhao
authors: Ya
authors: Y
authors: Li
authors: Xiao-Yan
authors: XY
authors: Liu
authors: Xian-Hua
authors: XH
authors: Chen
authors: Ping
authors: P
authors: Xu
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012694:Serine / Q000737:chemistry*; D000068103:Serine-Arginine Splicing Factors; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D009419:Nerve Tissue Proteins; D019913:Nuclear Localization Signals; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C097083:TRA2B protein, human; D000068103:Serine-Arginine Splicing Factors; D012694:Serine
references: J Neurosci. 2002 Jul 15;22(14):5889-99
references: 12122051
references: J Cell Biol. 2000 Apr 17;149(2):307-16
references: 10769024
references: Cell. 2007 Dec 14;131(6):1190-203
references: 18083107
references: Cell Biol Int. 2003;27(6):491-6
references: 12798777
references: PLoS Genet. 2011 Dec;7(12):e1002390
references: 22194695
references: J Cell Biol. 1997 Jul 28;138(2):225-38
references: 9230067
references: Cell. 2000 Oct 27;103(3):367-70
references: 11081623
references: J Neurotrauma. 2007 Apr;24(4):722-31
references: 17439354
references: Cancer Res. 2006 May 1;66(9):4774-80
references: 16651431
references: Mol Biol Cell. 1996 Oct;7(10):1559-72
references: 8898362
references: Genes Dev. 2004 Apr 1;18(7):755-68
references: 15082528
references: Cell. 1984 Dec;39(3 Pt 2):499-509
references: 6096007
references: J Biol Chem. 2003 May 16;278(20):18050-5
references: 12637531
references: Cell. 1993 Apr 9;73(1):47-59
references: 8462102
references: J Biol Chem. 2006 Feb 3;281(5):2460-9
references: 16308321
references: J Cell Biol. 1996 May;133(4):719-32
references: 8666659
references: J Cell Biol. 1994 Feb;124(3):249-60
references: 8294510
references: Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11524-8
references: 8524796
references: Mol Cell. 2008 Aug 8;31(3):438-48
references: 18691976
references: Genes Dev. 2002 Mar 1;16(5):594-607
references: 11877379
references: Genes Dev. 1998 Jan 1;12(1):55-66
references: 9420331
references: Trends Cell Biol. 1997 Apr;7(4):135-8
references: 17708924
references: J Cell Biol. 1998 Feb 23;140(4):737-50
references: 9472028
references: Hum Mol Genet. 2004 Mar 1;13(5):509-24
references: 14709600
references: Cell. 1991 Oct 18;67(2):335-42
references: 1655279
references: Trends Genet. 2001 Feb;17(2):100-7
references: 11173120
references: Genes Dev. 1992 May;6(5):837-47
references: 1577277
references: Genomics. 1998 Oct 15;53(2):191-202
references: 9790768
references: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9618-23
references: 10931943
references: Eur J Neurosci. 1999 Mar;11(3):788-802
references: 10103073
references: Nature. 1994 Jun 23;369(6482):678-82
references: 8208298
references: J Biol Chem. 2003 May 23;278(21):18997-9007
references: 12649279
references: Genes Dev. 1996 Jul 1;10(13):1569-79
references: 8682289
references: J Neurochem. 2006 Feb;96(3):635-44
references: 16371011
references: J Cell Biol. 1988 Sep;107(3):841-9
references: 3417784
references: EMBO J. 1996 Jan 15;15(2):265-75
references: 8617202
references: DNA Cell Biol. 1997 Jun;16(6):679-90
references: 9212162
references: Cell. 2006 Nov 3;127(3):635-48
references: 17081983
references: J Biol Chem. 2000 Mar 17;275(11):7950-7
references: 10713112
references: Mol Cell. 2003 Mar;11(3):837-43
references: 12667464
references: J Cell Biol. 1999 Jun 14;145(6):1145-52
references: 10366588
references: Anal Chem. 2008 Dec 15;80(24):9526-33
references: 19007248
references: Genes Dev. 2003 Feb 15;17(4):419-37
references: 12600935
references: RNA. 1995 Sep;1(7):663-80
references: 7585252
references: Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10154-9
references: 11517331
references: Proc Natl Acad Sci U S A. 2008 Aug 5;105(31):10762-7
references: 18669648
references: Annu Rev Cell Biol. 1993;9:265-315
references: 8280462
references: FEBS Lett. 1996 Jun 3;387(2-3):152-6
references: 8674539
references: Genes Dev. 1990 May;4(5):789-805
references: 2116360
references: Circ Res. 2008 Aug 29;103(5):485-92
references: 18669920