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The C-terminal domain of coilin interacts with Sm proteins and U snRNPs

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Abstract

Coilin is the signature protein of the Cajal body (CB), a nuclear suborganelle involved in the biogenesis of small nuclear ribonucleoproteins (snRNPs). Newly imported Sm-class snRNPs are thought to traffic through CBs before proceeding to their final nuclear destinations. Loss of coilin function in mice leads to significant viability and fertility problems. Coilin interacts directly with the spinal muscular atrophy (SMA) protein via dimethylarginine residues in its C-terminal domain. Although coilin hypomethylation results in delocalization of survival of motor neurons (SMN) from CBs, high concentrations of snRNPs remain within these structures. Thus, CBs appear to be involved in snRNP maturation, but factors that tether snRNPs to CBs have not been described. In this report, we demonstrate that the coilin C-terminal domain binds directly to various Sm and Lsm proteins via their Sm motifs. We show that the region of coilin responsible for this binding activity is separable from that which binds to SMN. Interestingly, U2, U4, U5, and U6 snRNPs interact with the coilin C-terminal domain in a glutathione S-transferase (GST)-pulldown assay, whereas U1 and U7 snRNPs do not. Thus, the ability to interact with free Sm (and Lsm) proteins as well as with intact snRNPs, indicates that coilin and CBs may facilitate the modification of newly formed snRNPs, the regeneration of ‘mature’ snRNPs, or the reclamation of unassembled snRNP components.

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References

  • Bauer DW, Gall JG (1997) Coiled bodies without coilin. Mol Biol Cell 8:73–82

    PubMed  CAS  Google Scholar 

  • Bell M, Schreiner S, Damianov A, Reddy R, Bindereif A (2002) p110, a novel human U6 snRNP protein and U4/U6 snRNP recycling factor. EMBO J 21:2724–2735

    Article  PubMed  CAS  Google Scholar 

  • Bellini M, Gall JG (1998) Coilin can form a complex with the U7 small nuclear ribonucleoprotein. Mol Biol Cell 9:2987–3001

    PubMed  CAS  Google Scholar 

  • Boisvert FM, Cote J, Boulanger MC, Cleroux P, Bachand F, Autexier C, Richard S (2002) Symmetrical dimethylarginine methylation is required for the localization of SMN in Cajal bodies and pre-mRNA splicing. J Cell Biol 159:957–969

    Article  PubMed  CAS  Google Scholar 

  • Brahms H, Meheus L, de Brabandere V, Fischer U, Luhrmann R (2001) Symmetrical dimethylation of arginine residues in spliceosomal Sm protein B/B′ and the Sm-like protein LSm4, and their interaction with the SMN protein. RNA 7:1531–1542

    Article  PubMed  CAS  Google Scholar 

  • Carmo-Fonseca M (2002a) The contribution of nuclear compartmentalization to gene regulation. Cell 108:513–521

    Article  PubMed  CAS  Google Scholar 

  • Carmo-Fonseca M (2002b) New clues to the function of the Cajal body. EMBO Rep 3:726–727

    Article  PubMed  CAS  Google Scholar 

  • Darzacq X, Jady BE, Verheggen C, Kiss AM, Bertrand E, Kiss T (2002) Cajal body-specific small nuclear RNAs: a novel class of 2′-O-methylation and pseudouridylation guide RNAs. EMBO J 21:2746–2756

    Article  PubMed  CAS  Google Scholar 

  • Dominski Z, Marzluff WF (1999) Formation of the 3′ end of histone mRNA. Gene 239:1–14

    Article  PubMed  CAS  Google Scholar 

  • Frey MR, Matera AG (1995) Coiled bodies contain U7 small nuclear RNA and associate with specific DNA sequences in interphase human cells. Proc Natl Acad Sci U S A 92:5915–5919

    Article  PubMed  CAS  Google Scholar 

  • Frey MR, Matera AG (2001) RNA-mediated interaction of Cajal bodies and U2 snRNA genes. J Cell Biol 154:499–509

    Article  PubMed  CAS  Google Scholar 

  • Frey MR, Bailey AD, Weiner AM, Matera AG (1999) Association of snRNA genes with coiled bodies is mediated by nascent snRNA transcripts. Curr Biol 9:126–135

    Article  PubMed  CAS  Google Scholar 

  • Friesen WJ, Dreyfuss G (2000) Specific sequences of the Sm and Sm-like (Lsm) proteins mediate their interaction with the spinal muscular atrophy disease gene product (SMN). J Biol Chem 275:26370–26375

    Article  PubMed  CAS  Google Scholar 

  • Friesen WJ, Massenet S, Paushkin S, Wyce A, Dreyfuss G (2001) SMN, the product of the spinal muscular atrophy gene, binds preferentially to dimethylarginine-containing protein targets. Mol Cell 7:1111–1117

    Article  PubMed  CAS  Google Scholar 

  • Gall JG (2000) Cajal bodies: the first 100 years. Annu Rev Cell Dev Biol 16:273–300

    Article  PubMed  CAS  Google Scholar 

  • Gall JG (2003) The centennial of the Cajal body. Nat Rev Mol Cell Biol 4:975–980

    Article  PubMed  CAS  Google Scholar 

  • Gall JG, Tsvetkov A, Wu Z, Murphy C (1995) Is the sphere organelle/coiled body a universal nuclear component? Dev Genet 16:25–35

    Article  PubMed  CAS  Google Scholar 

  • Gerbi SA, Borovjagin AV, Lange TS (2003) The nucleolus: a site of ribonucleoprotein maturation. Curr Opin Cell Biol 15:318–325

    Article  PubMed  CAS  Google Scholar 

  • He W, Parker R (2000) Functions of Lsm proteins in mRNA degradation and splicing. Curr Opin Cell Biol 12:346–350

    Article  PubMed  CAS  Google Scholar 

  • Hebert MD, Matera AG (2000) Self-association of coilin reveals a common theme in nuclear body localization. Mol Biol Cell 11:4159–4171

    PubMed  CAS  Google Scholar 

  • Hebert MD, Szymczyk PW, Shpargel KB, Matera AG (2001) Coilin forms the bridge between Cajal bodies and SMN, the spinal muscular atrophy protein. Genes Dev 15:2720–2729

    Article  PubMed  CAS  Google Scholar 

  • Hebert MD, Shpargel KB, Ospina JK, Tucker KE, Matera AG (2002) Coilin methylation regulates nuclear body formation. Dev Cell 3:329–337

    Article  PubMed  CAS  Google Scholar 

  • Huang Q, Pederson T (1999) A human U2 RNA mutant stalled in 3′ end processing is impaired in nuclear import. Nucleic Acids Res 27:1025–1031

    Article  PubMed  CAS  Google Scholar 

  • Isaac C, Yang Y, Meier UT (1998) Nopp140 functions as a molecular link between the nucleolus and the coiled bodies. J Cell Biol 142:319–329

    Article  PubMed  CAS  Google Scholar 

  • Jackson DA (2003) The principles of nuclear structure. Chromosome Res 11:387–401

    Article  PubMed  CAS  Google Scholar 

  • Jacobs EY, Frey MR, Wu W, Ingledue TC, Gebuhr TC, Gao L, Marzluff WF, Matera AG (1999) Coiled bodies preferentially associate with U4, U11, and U12 small nuclear RNA genes in interphase HeLa cells but not with U6 and U7 genes. Mol Biol Cell 10:1653–1663

    PubMed  CAS  Google Scholar 

  • Jady BE, Darzacq X, Tucker KE, Matera AG, Bertrand E, Kiss T (2003) Modification of Sm small nuclear RNAs occurs in the nucleoplasmic Cajal body following import from the cytoplasm. EMBO J 22:1878–1888

    Article  PubMed  CAS  Google Scholar 

  • Kambach C, Walke S, Nagai K (1999a) Structure and assembly of the spliceosomal small nuclear ribonucleoprotein particles. Curr Opin Struck Biol 9:222–230

    Article  CAS  Google Scholar 

  • Kambach C, Walke S, Young R, Avis JM, de la Fortelle E, Raker VA, Luhrmann R, Li J, Nagai K (1999b) Crystal structures of two Sm protein complexes and their implications for the assembly of the spliceosomal snRNPs. Cell 96:375–387

    Article  PubMed  CAS  Google Scholar 

  • Kiss T (2004) Biogenesis of small nuclear RNPs. J Cell Sci 117:5949–5951

    Article  PubMed  CAS  Google Scholar 

  • Lam YW, Lyon CE, Lamond AI (2002) Large-scale isolation of Cajal bodies from HeLa cells. Mol Biol Cell 13:2461–2473

    Article  PubMed  CAS  Google Scholar 

  • Lamond AI, Spector DL (2003) Nuclear speckles: a model for nuclear organelles. Nat Rev Mol Cell Biol 4:605–612

    Article  PubMed  CAS  Google Scholar 

  • Liu Q, Dreyfuss G (1996) A novel nuclear structure containing the survival of motor neurons protein. EMBO J 15:3555–3565

    PubMed  CAS  Google Scholar 

  • Massenet S, Pellizzoni L, Paushkin S, Mattaj IW, Dreyfuss G (2002) The SMN complex is associated with snRNPs throughout their cytoplasmic assembly pathway. Mol Cell Biol 22:6533–6541

    Article  PubMed  CAS  Google Scholar 

  • Matera AG (1999) RNA splicing: more clues from spinal muscular atrophy. Curr Biol 9:R140–R142

    Article  PubMed  CAS  Google Scholar 

  • Matera AG (2003) Cajal bodies. Curr Biol 13:R503

    Article  PubMed  CAS  Google Scholar 

  • Matera AG, Ward DC (1993) Nucleoplasmic organization of small nuclear ribonucleoproteins in cultured human cells. J Cell Biol 121:715–727

    Article  PubMed  CAS  Google Scholar 

  • Meister G, Fischer U (2002) Assisted RNP assembly: SMN and PRMT5 complexes cooperate in the formation of spliceosomal UsnRNPs. EMBO J 21:5853–5863

    Article  PubMed  CAS  Google Scholar 

  • Meister G, Buhler D, Pillai R, Lottspeich F, Fischer U (2001) A multiprotein complex mediates the ATP-dependent assembly of spliceosomal U snRNPs. Nat Cell Biol 3:945–949

    Article  PubMed  CAS  Google Scholar 

  • Meister G, Eggert C, Fischer U (2002) SMN-mediated assembly of RNPs: a complex story. Trends Cell Biol 12:472–478

    Article  PubMed  CAS  Google Scholar 

  • Mouaikel J, Narayanan U, Verheggen C, Matera AG, Bertrand E, Tazi J, Bordonne R (2003) Interaction between the small-nuclear-RNA cap hypermethylase and the spinal muscular atrophy protein, survival of motor neuron. EMBO Rep 4:616–622

    Article  PubMed  CAS  Google Scholar 

  • Muller B, Schumperli D (1997) The U7 snRNP and the hairpin binding protein: key players in histone mRNA metabolism. Semin Cell Dev Biol 8:567–576

    Article  PubMed  CAS  Google Scholar 

  • Narayanan U, Ospina JK, Frey MR, Hebert MD, Matera AG (2002) SMN, the spinal muscular atrophy protein, forms a pre-import snRNP complex with snurportin1 and importin beta. Hum Mol Genet 11:1785–1795

    Article  PubMed  CAS  Google Scholar 

  • Narayanan U, Achsel T, Lührmann R, Matera AG (2004) Coupled in vitro import of U snRNPs and SMN, the spinal muscular atrophy protein. Mol Cell 16:223–234

    Article  PubMed  CAS  Google Scholar 

  • Nesic D, Tanackovic G, Kramer A (2004) A role for Cajal bodies in the final steps of U2 snRNP biogenesis. J Cell Sci 117:4423–4433

    Article  PubMed  CAS  Google Scholar 

  • Niedojadlo J, Gorska-Brylass A (2003) New type of snRNP containing nuclear bodies in plant cells. Biol Cell 95:303–310

    Article  PubMed  CAS  Google Scholar 

  • Nottrott S, Urlaub H, Luhrmann R (2002) Hierarchical, clustered protein interactions with U4/U6 snRNA: a biochemical role for U4/U6 proteins. EMBO J 21:5527–5538

    Article  PubMed  CAS  Google Scholar 

  • Ogg SC, Lamond AI (2002) Cajal bodies and coilin-moving towards function. J Cell Biol 159:17–21

    Article  PubMed  CAS  Google Scholar 

  • Paushkin S, Gubitz AK, Massenet S, Dreyfuss G (2002) The SMN complex, an assembly some of ribonucleoproteins. Curr Opin Cell Biol 14:305–312

    Article  PubMed  CAS  Google Scholar 

  • Pellizzoni L, Kataoka N, Charroux B, Dreyfuss G (1998) A novel function for SMN, the spinal muscular atrophy disease gene product, in pre-mRNA splicing. Cell 95:615–624

    Article  PubMed  CAS  Google Scholar 

  • Pellizzoni L, Yong J, Dreyfuss G (2002) Essential role for the SMN complex in the specificity of snRNP assembly. Science 298:1775–1779

    Article  PubMed  CAS  Google Scholar 

  • Pillai RS, Will CL, Luhrmann R, Schumperli D, Muller B (2001) Purified U7 snRNPs lack the Sm proteins D1 and D2 but contain Lsm10, a new 14 kDa Sm D1-like protein. EMBO J 20:5470–5479

    Article  PubMed  CAS  Google Scholar 

  • Pillai RS, Grimmler M, Meister G, Will CL, Luhrmann R, Fischer U, Schumperli D (2003) Unique Sm core structure of U7 snRNPs: assembly by a specialized SMN complex and the role of a new component, Lsm11, in histone RNA processing. Genes Dev 17:2321–2333

    Article  PubMed  CAS  Google Scholar 

  • Plessel G, Luhrmann R, Kastner B (1997) Electron microscopy of assembly intermediates of the snRNP core: morphological similarities between the RNA-free (E.F.G) protein heteromer and the intact snRNP core. J Mol Biol 265:87–94

    Article  PubMed  CAS  Google Scholar 

  • Raker VA, Plessel G, Luhrmann R (1996) The snRNP core assembly pathway: identification of stable core protein heteromeric complexes and an snRNP subcore particle in vitro. EMBO J 15:2256–2269

    PubMed  CAS  Google Scholar 

  • Schul W, van Driel R, de Jong L (1998) Coiled bodies and U2 snRNA genes adjacent to coiled bodies are enriched in factors required for snRNA transcription. Mol Biol Cell 9:1025–1036

    PubMed  CAS  Google Scholar 

  • Schumperli D, Pillai RS (2004) The special Sm core structure of the U7 snRNP: far-reaching significance of a small nuclear ribonucleoprotein. Cell Mol Life Sci 60:in press

  • Selenko P, Sprangers R, Stier G, Buhler D, Fischer U, Sattler M (2001) SMN tudor domain structure and its interaction with the Sm proteins. Nat Struct Biol 8:27–31

    Article  PubMed  CAS  Google Scholar 

  • Sleeman JE, Lamond AI (1999) Newly assembled snRNPs associate with coiled bodies before speckles, suggesting a nuclear snRNP maturation pathway. Curr Biol 9:1065–1074

    Article  PubMed  CAS  Google Scholar 

  • Smith KP, Carter KC, Johnson CV, Lawrence JB (1995) U2 and U1 snRNA gene loci associate with coiled bodies. J Cell Biochem 59:473–485

    Article  PubMed  CAS  Google Scholar 

  • Spector DL (2003) The dynamics of chromosome organization and gene regulation. Annu Rev Biochem 72:573–608

    Article  PubMed  CAS  Google Scholar 

  • Stanek D, Neugebauer KM (2004) Detection of snRNP assembly intermediates in Cajal bodies by fluorescence resonance energy transfer. J Cell Biol 166:1015–1025

    Article  PubMed  CAS  Google Scholar 

  • Stanek D, Rader SD, Klingauf M, Neugebauer KM (2003) Targeting of U4/U6 small nuclear RNP assembly factor SART3/p110 to Cajal bodies. J Cell Biol 160:505–516

    Article  PubMed  CAS  Google Scholar 

  • Stefanovic B, Hackl W, Luhrmann R, Schumperli D (1995) Assembly, nuclear import and function of U7 snRNPs studied by microinjection of synthetic U7 RNA into Xenopus oocytes. Nucleic Acids Res 23:3141–3151

    Article  PubMed  CAS  Google Scholar 

  • Sumpter V, Kahrs A, Fischer U, Kornstadt U, Luhrmann R (1992) In vitro reconstitution of U1 and U2 snRNPs from isolated proteins and snRNA. Mol Biol Rep 16:229–240

    Article  PubMed  CAS  Google Scholar 

  • Tanackovic G, Kramer A (2005) Human splicing factor SF3a, but not SF1, is essential for pre-mRNA splicing in vivo. Mol Biol Cell 16:1366–1377

    Article  PubMed  CAS  Google Scholar 

  • Tucker KE, Matera AG (2004) The Cajal body: a nuclear gathering place. In: Hemmerich P, Diekmann S (eds) Visions of the nucleus. American Scientific Publishers, New York, NY

    Google Scholar 

  • Tucker KE, Berciano MT, Jacobs EY, LePage DF, Shpargel KB, Rossire JJ, Chan EK, Lafarga M, Conlon RA, Matera AG (2001) Residual Cajal bodies in coilin knockout mice fail to recruit Sm snRNPs and SMN, the spinal muscular atrophy gene product. J Cell Biol 154:293–307

    Article  PubMed  CAS  Google Scholar 

  • Verheggen C, Lafontaine DL, Samarsky D, Mouaikel J, Blanchard JM, Bordonne R, Bertrand E (2002) Mammalian and yeast U3 snoRNPs are matured in specific and related nuclear compartments. EMBO J 21:2736–2745

    Article  PubMed  CAS  Google Scholar 

  • Will CL, Luhrmann R (2001) Spliceosomal UsnRNP biogenesis, structure and function. Curr Opin Cell Biol 13:290–301

    Article  PubMed  CAS  Google Scholar 

  • Wolin SL, Cedervall T (2002) The La protein. Annu Rev Biochem 71:375–403

    Article  PubMed  CAS  Google Scholar 

  • Yannoni YM, White K (1997) Association of the neuron-specific RNA binding domain-containing protein ELAV with the coiled body in Drosophila neurons. Chromosoma 105:332–341

    Article  PubMed  CAS  Google Scholar 

  • Yong J, Golembe TJ, Battle DJ, Pellizzoni L, Dreyfuss G (2004) snRNAs contain specific SMN-binding domains that are essential for snRNP assembly. Mol Cell Biol 24:2747–2756

    Article  PubMed  CAS  Google Scholar 

  • Yu YT, Shu MD, Steitz JA (1998) Modifications of U2 snRNA are required for snRNP assembly and pre-mRNA splicing. EMBO J 17:5783–5795

    Article  PubMed  CAS  Google Scholar 

  • Zaric B, Chami M, Remigy H, Engel A, Ballmer-Hofer K, Winkler FK, Kambach C (2005) Reconstitution of two recombinant LSm protein complexes reveals aspects of their architecture, assembly, and function. JBC 280:16066–16075

    Article  CAS  Google Scholar 

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Acknowledgements

We thank R. Lührmann, C.L. Will, and H. Salz for providing us with reagents used in this work. Moreover, we acknowledge financial support by the Novartis Foundation, the State of Bern, the Swiss National Science Foundation, the Muscular Dystrophy Association of the USA, and the US National Institutes of Health (R01 grants GM53034 and NS41617).

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  1. Ramesh S. Pillai

    Present address: Friedrich Miescher Institute, Maulbeerstrasse 66, 4058, Basel, Switzerland

Authors and Affiliations

  1. Department of Biochemistry, The University of Mississippi Medical Center, Jackson, MS, 39216-4505, USA

    Hongzhi Xu & Michael D. Hebert

  2. Institute of Cell Biology, University of Bern, Baltzerstrasse 4, 3012, Bern, Switzerland

    Ramesh S. Pillai, Teldja N. Azzouz & Daniel Schümperli

  3. Department of Genetics, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH, 44106-4955, USA

    Karl B. Shpargel, Michael D. Hebert & A. Gregory Matera

  4. Paul Scherrer Institute, Life Sciences, 5232, Villigen PSI, Switzerland

    Christian Kambach

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Xu, H., Pillai, R.S., Azzouz, T.N. et al. The C-terminal domain of coilin interacts with Sm proteins and U snRNPs. Chromosoma 114, 155–166 (2005). https://doi.org/10.1007/s00412-005-0003-y

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