Home LiteratureArticle Details
PMID: 20237019 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Dynamic nucleocytoplasmic shuttling of an Arabidopsis SR splicing factor: role of the RNA-binding domains.

Plant physiology ·Vol. 153 ·No. 1 ·2010-05-00 ·Pages 273-84

Rausin G, Tillemans V, Stankovic N, Hanikenne M, Motte P

Abstract

Serine/arginine-rich (SR) proteins are essential nuclear-localized splicing factors. We have investigated the dynamic subcellular distribution of the Arabidopsis (Arabidopsis thaliana) RSZp22 protein, a homolog of the human 9G8 SR factor. Little is known about the determinants underlying the control of plant SR protein dynamics, and so far most studies relied on ectopic transient overexpression. Here, we provide a detailed analysis of the RSZp22 expression profile and describe its nucleocytoplasmic shuttling properties in specific cell types. Comparison of transient ectopic- and stable tissue-specific expression highlights the advantages of both approaches for nuclear protein dynamic studies. By site-directed mutagenesis of RSZp22 RNA-binding sequences, we show that functional RNA recognition motif RNP1 and zinc-knuckle are dispensable for the exclusive protein nuclear localization and speckle-like distribution. Fluorescence resonance energy transfer imaging also revealed that these motifs are implicated in RSZp22 molecular interactions. Furthermore, the RNA-binding motif mutants are defective for their export through the CRM1/XPO1/Exportin-1 receptor pathway but retain nucleocytoplasmic mobility. Moreover, our data suggest that CRM1 is a putative export receptor for mRNPs in plants.

MeSH Terms
Active Transport, Cell Nucleus Arabidopsis/metabolism Arabidopsis Proteins/metabolism Cell Nucleus/metabolism Genes, Reporter Mutagenesis, Site-Directed Protein Interaction Domains and Motifs RNA-Binding Proteins/metabolism Recombinant Fusion Proteins/metabolism Serine-Arginine Splicing Factors
Chemicals
Arabidopsis Proteins RNA-Binding Proteins RSZ protein, Arabidopsis Recombinant Fusion Proteins Serine-Arginine Splicing Factors
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Rausin Glwadys
Laboratory of Functional Genomics and Plant Molecular Imaging, Department of Life Sciences, Institute of Botany, University of Liège, B-4000 Liege, Belgium.
Tillemans Vinciane
Stankovic Nancy
Hanikenne Marc
Motte Patrick
References (68)
68 references, click to expand
  1. RNA recognition by RNP proteins during RNA processing.
    Annu Rev Biophys Biomol Struct. 1998;27:407-45 PMID: 9646873
  2. The splicing factors 9G8 and SRp20 transactivate splicing through different and specific enhancers.
    RNA. 1999 Mar;5(3):468-83 PMID: 10094314
  3. A novel family of plant splicing factors with a Zn knuckle motif: examination of RNA binding and splicing activities.
    Plant Mol Biol. 1999 Mar;39(4):761-73 PMID: 10350090
  4. Involvement of SR proteins in mRNA surveillance.
    Mol Cell. 2004 Nov 19;16(4):597-607 PMID: 15546619
  5. Early flower development in Arabidopsis.
    Plant Cell. 1990 Aug;2(8):755-67 PMID: 2152125
  6. Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains.
    EMBO J. 1993 Dec;12(12):4715-26 PMID: 8223480
  7. The dynamics of a pre-mRNA splicing factor in living cells.
    Nature. 1997 May 29;387(6632):523-7 PMID: 9168118
  8. Role of the modular domains of SR proteins in subnuclear localization and alternative splicing specificity.
    J Cell Biol. 1997 Jul 28;138(2):225-38 PMID: 9230067
  9. qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data.
    Genome Biol. 2007;8(2):R19 PMID: 17291332
  10. A regulatory role for CRM1 in the multi-directional trafficking of splicing snRNPs in the mammalian nucleus.
    J Cell Sci. 2007 May 1;120(Pt 9):1540-50 PMID: 17405816
  11. Alternative splicing of pre-messenger RNAs in plants in the genomic era.
    Annu Rev Plant Biol. 2007;58:267-94 PMID: 17222076
  12. SR proteins function in coupling RNAP II transcription to pre-mRNA splicing.
    Mol Cell. 2007 Jun 22;26(6):867-81 PMID: 17588520
  13. The maximal size of protein to diffuse through the nuclear pore is larger than 60kDa.
    FEBS Lett. 2007 Jul 10;581(17):3164-70 PMID: 17588566
  14. The shuttling SR protein 9G8 plays a role in translation of unspliced mRNA containing a constitutive transport element.
    J Biol Chem. 2007 Jul 6;282(27):19844-53 PMID: 17513303
  15. Structural and functional analysis of RNA and TAP binding to SF2/ASF.
    EMBO Rep. 2007 Aug;8(8):756-62 PMID: 17668007
  16. The nuclear pore comes to the fore.
    Trends Plant Sci. 2008 Jan;13(1):20-7 PMID: 18155634
  17. Analyses of in vivo interaction and mobility of two spliceosomal proteins using FRAP and BiFC.
    PLoS One. 2008;3(4):e1953 PMID: 18414657
  18. The splicing factor SF2/ASF regulates translation initiation by enhancing phosphorylation of 4E-BP1.
    Mol Cell. 2008 Apr 25;30(2):179-89 PMID: 18439897
  19. Eleven golden rules of quantitative RT-PCR.
    Plant Cell. 2008 Jul;20(7):1736-7 PMID: 18664613
  20. Towards a systematic validation of references in real-time rt-PCR.
    Plant Cell. 2008 Jul;20(7):1734-5 PMID: 18664615
  21. Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei.
    Exp Cell Res. 2008 Oct 15;314(17):3175-86 PMID: 18674533
  22. The SR protein family of splicing factors: master regulators of gene expression.
    Biochem J. 2009 Jan 1;417(1):15-27 PMID: 19061484
  23. The spliceosome: design principles of a dynamic RNP machine.
    Cell. 2009 Feb 20;136(4):701-18 PMID: 19239890
  24. Proteomic analysis of the Arabidopsis nucleolus suggests novel nucleolar functions.
    Mol Biol Cell. 2005 Jan;16(1):260-9 PMID: 15496452
  25. Functional distribution and dynamics of Arabidopsis SR splicing factors in living plant cells.
    Plant J. 2005 Feb;41(4):567-82 PMID: 15686520
  26. Understanding alternative splicing: towards a cellular code.
    Nat Rev Mol Cell Biol. 2005 May;6(5):386-98 PMID: 15956978
  27. Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis.
    Plant Physiol. 2005 Sep;139(1):5-17 PMID: 16166256
  28. Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light.
    Nat Biotechnol. 2006 Apr;24(4):461-5 PMID: 16550175
  29. Zinc-dependent global transcriptional control, transcriptional deregulation, and higher gene copy number for genes in metal homeostasis of the hyperaccumulator Arabidopsis halleri.
    Plant Physiol. 2006 Sep;142(1):148-67 PMID: 16844841
  30. Molecular basis of RNA recognition and TAP binding by the SR proteins SRp20 and 9G8.
    EMBO J. 2006 Nov 1;25(21):5126-37 PMID: 17036044
  31. Insights into nuclear organization in plants as revealed by the dynamic distribution of Arabidopsis SR splicing factors.
    Plant Cell. 2006 Nov;18(11):3218-34 PMID: 17114353
  32. Biochemical and cellular characterization of the plant ortholog of PYM, a protein that interacts with the exon junction complex core proteins Mago and Y14.
    Planta. 2007 Feb;225(3):625-39 PMID: 16953428
  33. SDE5, the putative homologue of a human mRNA export factor, is required for transgene silencing and accumulation of trans-acting endogenous siRNA.
    Plant J. 2007 Apr;50(1):140-8 PMID: 17397509
  34. CRM1-mediated nuclear export: to the pore and beyond.
    Trends Cell Biol. 2007 Apr;17(4):193-201 PMID: 17317185
  35. A plethora of plant serine/arginine-rich proteins: redundancy or evolution of novel gene functions?
    Biochem Soc Trans. 2004 Aug;32(Pt 4):561-4 PMID: 15270675
  36. Splicing factor hSlu7 contains a unique functional domain required to retain the protein within the nucleus.
    Mol Biol Cell. 2004 Aug;15(8):3782-95 PMID: 15181151
  37. Uncapped mRNA introduced into tobacco protoplasts can be imported into the nucleus and is trapped by leptomycin B.
    Plant Cell Rep. 2004 Aug;23(1-2):99-103 PMID: 15221275
  38. A compendium of methods useful for characterizing Arabidopsis pollen mutants and gametophytically-expressed genes.
    Plant J. 2004 Sep;39(5):761-75 PMID: 15315637
  39. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.
    EMBO J. 1987 Dec 20;6(13):3901-7 PMID: 3327686
  40. SR protein family members display diverse activities in the formation of nascent and mature mRNPs in vivo.
    Mol Cell. 2009 Apr 24;34(2):179-90 PMID: 19394295
  41. mRNA nuclear export at a glance.
    J Cell Sci. 2009 Jun 15;122(Pt 12):1933-7 PMID: 19494120
  42. SR proteins in vertical integration of gene expression from transcription to RNA processing to translation.
    Mol Cell. 2009 Jul 10;35(1):1-10 PMID: 19595711
  43. Dynamic behavior of Arabidopsis eIF4A-III, putative core protein of exon junction complex: fast relocation to nucleolus and splicing speckles under hypoxia.
    Plant Cell. 2009 May;21(5):1592-606 PMID: 19435936
  44. Aberrant mRNA transcripts and the nonsense-mediated decay proteins UPF2 and UPF3 are enriched in the Arabidopsis nucleolus.
    Plant Cell. 2009 Jul;21(7):2045-57 PMID: 19602621
  45. Mechanisms of alternative splicing regulation: insights from molecular and genomics approaches.
    Nat Rev Mol Cell Biol. 2009 Nov;10(11):741-54 PMID: 19773805
  46. A human importin-beta family protein, transportin-SR2, interacts with the phosphorylated RS domain of SR proteins.
    J Biol Chem. 2000 Mar 17;275(11):7950-7 PMID: 10713112
  47. Sorting out the complexity of SR protein functions.
    RNA. 2000 Sep;6(9):1197-211 PMID: 10999598
  48. The balance of nuclear import and export determines the intracellular distribution and function of tomato heat stress transcription factor HsfA2.
    Mol Cell Biol. 2001 Mar;21(5):1759-68 PMID: 11238913
  49. Nucleocytoplasmic shuttling of heterodimeric splicing factor U2AF.
    J Biol Chem. 2001 Apr 20;276(16):13104-12 PMID: 11118443
  50. Splicing factors SRp20 and 9G8 promote the nucleocytoplasmic export of mRNA.
    Mol Cell. 2001 Apr;7(4):899-905 PMID: 11336712
  51. Delineation of mRNA export pathways by the use of cell-permeable peptides.
    Science. 2001 Nov 30;294(5548):1895-901 PMID: 11729309
  52. Directed proteomic analysis of the human nucleolus.
    Curr Biol. 2002 Jan 8;12(1):1-11 PMID: 11790298
  53. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes.
    Genome Biol. 2002 Jun 18;3(7):RESEARCH0034 PMID: 12184808
  54. Nuclear export and retention signals in the RS domain of SR proteins.
    Mol Cell Biol. 2002 Oct;22(19):6871-82 PMID: 12215544
  55. Network of interactions of a novel plant-specific Arg/Ser-rich protein, atRSZ33, with atSC35-like splicing factors.
    J Biol Chem. 2002 Oct 18;277(42):39989-98 PMID: 12176998
  56. Functional proteomic analysis of human nucleolus.
    Mol Biol Cell. 2002 Nov;13(11):4100-9 PMID: 12429849
  57. Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data.
    Neurosci Lett. 2003 Mar 13;339(1):62-6 PMID: 12618301
  58. SR splicing factors serve as adapter proteins for TAP-dependent mRNA export.
    Mol Cell. 2003 Mar;11(3):837-43 PMID: 12667464
  59. Regulation of adenovirus alternative RNA splicing correlates with a reorganization of splicing factors in the nucleus.
    Exp Cell Res. 2003 Sep 10;289(1):77-85 PMID: 12941606
  60. Nucleo-cytoplasmic partitioning of proteins in plants: implications for the regulation of environmental and developmental signalling.
    Curr Genet. 2003 Dec;44(5):231-60 PMID: 14523572
  61. Arginine-serine-rich domains bound at splicing enhancers contact the branchpoint to promote prespliceosome assembly.
    Mol Cell. 2004 Feb 13;13(3):367-76 PMID: 14967144
  62. Nuclear bodies and compartmentalization of pre-mRNA splicing factors in higher plants.
    Chromosoma. 2004 Feb;112(5):255-66 PMID: 14740228
  63. CRM1-dependent, but not ARE-mediated, nuclear export of IFN-alpha1 mRNA.
    J Cell Sci. 2004 May 1;117(Pt 11):2259-70 PMID: 15126627
  64. Tissue-specific expression and dynamic organization of SR splicing factors in Arabidopsis.
    Mol Biol Cell. 2004 Jun;15(6):2664-73 PMID: 15034145
  65. Characterization of C-terminal domains of Arabidopsis heat stress transcription factors (Hsfs) and identification of a new signature combination of plant class A Hsfs with AHA and NES motifs essential for activator function and intracellular localization.
    Plant J. 2004 Jul;39(1):98-112 PMID: 15200645
  66. Broad specificity of SR (serine/arginine) proteins in the regulation of alternative splicing of pre-messenger RNA.
    Prog Nucleic Acid Res Mol Biol. 2004;78:37-88 PMID: 15210328
  67. Hypophosphorylated ASF/SF2 binds TAP and is present in messenger ribonucleoproteins.
    J Biol Chem. 2004 Jul 23;279(30):31745-9 PMID: 15184380
  68. A specific subset of SR proteins shuttles continuously between the nucleus and the cytoplasm.
    Genes Dev. 1998 Jan 1;12(1):55-66 PMID: 9420331
Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
1532-2548
DOI
10.1104/pp.110.154740
Published
2010-05-00
Epub
2010-00-17
Pages
273-84
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC2862426
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.110.154740 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2