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Caenorhabditis elegans in the study of SMN-interacting proteins: a role for SMI-1, an orthologue of human Gemin2 and the identification of novel components of the SMN complex

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Invertebrate Neuroscience

Abstract

Spinal muscular atrophy is a common neuromuscular disorder caused by mutations in the survival motor neuron (SMN) gene. In mammals, SMN is tightly associated with Gemin2. To gain further insight into the functions of SMN and Gemin2, we have cloned and sequenced smi-1 (Survival of Motor neuron-Interacting protein 1), a C. elegans homologue of the human Gemin2 gene. We show that the SMI-1 expression pattern and RNA interference phenotype show considerable overlap with that previously reported for SMN-1. Finally, we demonstrate that the SMN-1 and SMI-1 proteins directly interact. Having demonstrated the utility of the C. elegans genetic model for investigating genes encoding SMN-interacting proteins, we have undertaken a yeast two-hybrid screen of a C. elegans cDNA library to identify novel proteins that interact with SMN-1. We show the direct interaction of SMN-1 with nine novel proteins, several of which may be involved in RNA metabolism.

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References

  • Aerbajinai W, Ishihara T, Arahata K, Tsukahara T (2002) Increased expression level of the splicing variant of SIP1 in motor neuron diseases. Int J Biochem Cell Biol 34:699–707

    Article  PubMed  CAS  Google Scholar 

  • Ahringer J (1997) Turn to the worm! Curr Opin Genet Dev 7:410–415

    Article  PubMed  CAS  Google Scholar 

  • Alen C, Kent NA, Jones HS, O’Sullivan J, Aranda A, Proudfoot NJ (2002) A role for chromatin remodeling in transcriptional termination by RNA polymerase II. Mol Cell 10:1441–52

    Article  PubMed  CAS  Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Article  PubMed  CAS  Google Scholar 

  • Askjaer P, Galy V, Hannak E, Mattaj IW (2002) Ran GTPase cycle and importins alpha and beta are essential for spindle formation and nuclear envelope assembly in living Caenorhabditis elegans embryos. Mol Biol Cell 13:4355–4370

    Article  PubMed  CAS  Google Scholar 

  • Auerbach D, Thaminy S, Hottiger MO, Stagljar I (2002) The post-genomic era of interactive proteomics: facts and perspectives. Proteomics 2:611–623

    Article  PubMed  CAS  Google Scholar 

  • Bargmann CI (1998) Neurobiology of the Caenorhabditis elegans genome. Science 282:2028–2033

    Article  PubMed  CAS  Google Scholar 

  • Bargmann C (2000) Simple organisms. Neurobiol Dis 7:520–522

    Article  PubMed  CAS  Google Scholar 

  • Barstead RJ, Waterston RH (1989) The basal component of the nematode dense-body is vinculin. J Biol Chem 264: 10177–10185

    PubMed  CAS  Google Scholar 

  • Brenner S (1974) The genetics of Caenorhabditis elegans. Genetics 77:71–94

    PubMed  CAS  Google Scholar 

  • Campbell L, Hunter K. M., Mohaghegh P, Tinsley JM, Brasch MA, Davies KE (2000) Direct interaction of Smn with dp103, a putative RNA helicase: a role for Smn in transcription regulation? Hum Mol Genet 9:1093–100

    Article  PubMed  CAS  Google Scholar 

  • The C. elegans Sequencing Consortium (1998) Genome sequence of the nematode C. elegans: a platform for investigating biology. Science 282:2012–2018

    Article  Google Scholar 

  • Charroux B, Pellizzoni L, Perkinson RA, Shevchenko A, Mann M, Dreyfuss G (1999) Gemin3: a novel DEAD box protein that interacts with SMN, the spinal muscular atrophy gene product, and is a component of gems. J Cell Biol 147:1181–1194

    Article  PubMed  CAS  Google Scholar 

  • Coovert DD, Le TT, McAndrew PE, Strasswimmer J, Crawford TO, Mendell JR, Coulson SE, Androphy EJ, Prior TW, Burghes AH (1997) The survival motor neuron protein in spinal muscular atrophy. Hum Mol Genet 6:1205–1214

    Article  PubMed  CAS  Google Scholar 

  • de la Cruz J, Kressler D, Linder P (1999) Unwinding RNA in Saccharomyces cerevisiae: DEAD-box proteins and related families. Trends Biochem Sci 24:192–198

    Article  Google Scholar 

  • Culetto E, Sattelle DB (2000) A role for Caenorhabditis elegans in understanding the function and interactions of human disease genes. Hum Mol Genet 9:869–877

    Article  PubMed  CAS  Google Scholar 

  • Dubowitz V (1995) Chaos in the classification of SMA: a possible resolution. Neuromuscul Disord 5:3–5

    Article  PubMed  CAS  Google Scholar 

  • Elledge SJ, Mulligan JT, Ramer SW, Spottswood M, Davis RW (1991) Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. Proc Natl Acad Sci USA 88:1731–1735

    Article  PubMed  CAS  Google Scholar 

  • Emery AE (1991) Population frequencies of inherited neuromuscular diseases—a world survey. Neuromuscul Disord 1:19–29

    Article  PubMed  CAS  Google Scholar 

  • Fire A, Harrison SW, Dixon D (1990) A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans. Gene 93:189–198

    Article  PubMed  CAS  Google Scholar 

  • Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811

    Article  PubMed  CAS  Google Scholar 

  • Fischer U, Liu Q, Dreyfuss G (1997) The SMN-SIP1 complex has an essential role in spliceosomal snRNP biogenesis. Cell 90:1023–1029

    Article  PubMed  CAS  Google Scholar 

  • Fraser AG, Kamath RS, Zipperlen P, Martinez-Campos M, Sohrmann M, Ahringer J (2000) Functional genomic analysis of C. elegans chromosome I by systematic RNA interference. Nature 408:325–330

    Article  PubMed  CAS  Google Scholar 

  • Gallegos M, Ahringer J, Crittenden S., Kimble J (1998) Repression by the 3′ UTR of fem-3, a sex-determining gene, relies on a ubiquitous mog-dependent control in Caenorhabditis elegans. Embo J 17:6337–6347

    Article  PubMed  CAS  Google Scholar 

  • Halle JP, Meisterernst M (1996) Gene expression: increasing evidence for a transcriptosome. Trends Genet 12:161–163

    Article  PubMed  CAS  Google Scholar 

  • Heaton JH, Dlakic WM, Dlakic M., Gelehrter TD (2001) Identification and cDNA cloning of a novel RNA-binding protein that interacts with the cyclic nucleotide-responsive sequence in the Type-1 plasminogen activator inhibitor mRNA. J Biol Chem 276:3341–3347

    Article  PubMed  CAS  Google Scholar 

  • Helmken C, Wetter A, Rudnik-Schoneborn S, Liehr T, Zerres K, Wirth B (2000) An essential SMN interacting protein (SIP1) is not involved in the phenotypic variability of spinal muscular atrophy (SMA). Eur J Hum Genet 8:493–499

    Article  PubMed  CAS  Google Scholar 

  • Hirose Y, Manley JL (2000) RNA polymerase II and the integration of nuclear events. Genes Dev 14:1415–1429

    PubMed  CAS  Google Scholar 

  • Hobert O, Moerman DG, Clark KA, Beckerle MC, Ruvkun G (1999) A conserved LIM protein that affects muscular adherens junction integrity and mechanosensory function in Caenorhabditis elegans. J Cell Biol 144:45–57

    Article  PubMed  CAS  Google Scholar 

  • Hodgkin J., Herman RK (1998) Changing styles in C. elegans genetics. Trends Genet 14:352–7

    Article  PubMed  CAS  Google Scholar 

  • Huang L, Grammatikakis N, Yoneda M, Banerjee SD, Toole BP (2000) Molecular characterization of a novel intracellular hyaluronan-binding protein. J Biol Chem 275:29829–29839

    Article  PubMed  CAS  Google Scholar 

  • Jablonka S, Schrank B, Kralewski M, Rossoll W, Sendtner M (2000) Reduced survival motor neuron (Smn) gene dose in mice leads to motor neuron degeneration: an animal model for spinal muscular atrophy type III. Hum Mol Genet 9:341–346

    Article  PubMed  CAS  Google Scholar 

  • Jablonka S, Bandilla M, Wiese S, Buhler D, Wirth B, Sendtner M, Fischer U (2001) Co-regulation of survival of motor neuron (SMN) protein and its interactor SIP1 during development and in spinal muscular atrophy. Hum Mol Genet 10:497–505

    Article  PubMed  CAS  Google Scholar 

  • Jablonka S, Holtmann B, Meister G, Bandilla M, Rossoll W, Fischer U, Sendtner M (2002) Gene targeting of Gemin2 in mice reveals a correlation between defects in the biogenesis of U snRNPs and motoneuron cell death. Proc Natl Acad Sci USA 99:10126–10131

    Article  PubMed  CAS  Google Scholar 

  • Johnstone IL, Barry JD (1996) Temporal reiteration of a precise gene expression pattern during nematode development. Embo J 15:3633–3639

    PubMed  CAS  Google Scholar 

  • Jones KW, Gorzynski K, Hales CM, Fischer U, Badbanchi F, Terns RM, Terns MP (2001) Direct interaction of the spinal muscular atrophy disease protein SMN with the small nucleolar RNA-associated protein fibrillarin. J Biol Chem 276:38645–38651

    Article  PubMed  CAS  Google Scholar 

  • Kamath RS, Fraser AG, Dong Y, Poulin G, Durbin R, Gotta M, Kanapin A, Le Bot N, Moreno S, Sohrmann M et al (2003) Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Nature 421:231–237

    Article  PubMed  CAS  Google Scholar 

  • Kramer JM, French RP, Park EC, Johnson JJ (1990) The Caenorhabditis elegans rol-6 gene, which interacts with the sqt-1 collagen gene to determine organismal morphology, encodes a collagen. Mol Cell Biol 10:2081–2089

    PubMed  CAS  Google Scholar 

  • Lefebvre S, Burglen L, Reboullet S, Clermont O, Burlet P, Viollet L, Benichou B, Cruaud C, Millasseau P, Zeviani M et al (1995) Identification and characterization of a spinal muscular atrophy-determining gene. Cell 80:155–165

    Article  PubMed  CAS  Google Scholar 

  • Lefebvre S, Burlet P, Liu Q, Bertrandy S, Clermont O, Munnich A, Dreyfuss G, Melki J (1997) Correlation between severity and SMN protein level in spinal muscular atrophy. Nat Genet 16:265–269

    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 

  • Liu Q, Fischer U, Wang F, Dreyfuss G (1997) The spinal muscular atrophy disease gene product, SMN, and its associated protein SIP1 are in a complex with spliceosomal snRNP proteins. Cell 90:1013–1021

    Article  PubMed  CAS  Google Scholar 

  • Lorson CL, Hahnen E, Androphy EJ, Wirth B (1999) A single nucleotide in the SMN gene regulates splicing and is responsible for spinal muscular atrophy. Proc Natl Acad Sci USA 96:6307–6311

    Article  PubMed  CAS  Google Scholar 

  • MacKenzie AE, Gendron NH (2001) Tudor reign. Nat Struct Biol 8:13–15

    Article  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 

  • Mello C, Fire A (1995) DNA transformation. Methods Cell Biol 48:451–482

    Article  PubMed  CAS  Google Scholar 

  • Miguel-Aliaga I, Culetto E, Walker DS, Baylis HA, Sattelle DB, Davies KE (1999) The Caenorhabditis elegans orthologue of the human gene responsible for spinal muscular atrophy is a maternal product critical for germline maturation and embryonic viability. Hum Mol Genet 8:2133–2143

    Article  PubMed  CAS  Google Scholar 

  • Monani UR, Lorson CL, Parsons DW, Prior TW, Androphy EJ, Burghes AH, McPherson JD (1999) A single nucleotide difference that alters splicing patterns distinguishes the SMA gene SMN1 from the copy gene SMN2. Hum Mol Genet 8:1177–1183

    Article  PubMed  CAS  Google Scholar 

  • Mourelatos Z, Abel L, Yong J, Kataoka N, Dreyfuss G (2001) SMN interacts with a novel family of hnRNP and spliceosomal proteins. Embo J 20:5443–52

    Article  PubMed  CAS  Google Scholar 

  • Neubauer G, King A, Rappsilber J, Calvio C, Watson M, Ajuh P, Sleeman J, Lamond A, Mann M (1998) Mass spectrometry and EST-database searching allows characterization of the multi-protein spliceosome complex. Nat Genet 20:46–50

    Article  PubMed  CAS  Google Scholar 

  • Park JW, Voss PG, Grabski S, Wang JL, Patterson RJ (2001) Association of galectin-1 and galectin-3 with Gemin4 in complexes containing the SMN protein. Nucleic Acids Res 29:3595–602

    Article  PubMed  CAS  Google Scholar 

  • Pearn J (1980) Classification of spinal muscular atrophies. Lancet 1:919–22

    Article  PubMed  CAS  Google Scholar 

  • Pellizzoni L, Charroux B, Rappsilber J, Mann M., Dreyfuss G (2001) A functional interaction between the survival motor neuron complex and RNA polymerase II. J Cell Biol 152:75–85

    Article  PubMed  CAS  Google Scholar 

  • Puoti A, Kimble J (2000) The hermaphrodite sperm/oocyte switch requires the Caenorhabditis elegans homologs of PRP2 and PRP22. Proc Natl Acad Sci USA 97:3276–3281

    Article  PubMed  CAS  Google Scholar 

  • Puoti A, Pugnale P, Belfiore M, Schlappi AC, Saudan Z (2001) RNA and sex determination in Caenorhabditis elegans. Post-transcriptional regulation of the sex-determining tra-2 and fem-3 mRNAs in the Caenorhabditis elegans hermaphrodite. EMBO Rep 2:899–904

    Article  PubMed  CAS  Google Scholar 

  • Ruvkun G, Hobert O (1998) The taxonomy of developmental control in Caenorhabditis elegans. Science 282:2033–2041

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russell D (1991) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbour

    Google Scholar 

  • Schultz J, Copley RR, Doerks T, Ponting CP, Bork P (2000) SMART: a web-based tool for the study of genetically mobile domains. Nucleic Acids Res 28:231–234

    Article  PubMed  CAS  Google Scholar 

  • Sonnichsen B, Koski LB, Walsh A, Marschall P, Neumann B, Brehm M, Alleaume AM, Artelt J, Bettencourt P, Cassin E et al (2005) Full-genome RNAi profiling of early embryogenesis in Caenorhabditis elegans. Nature 434:462–9

    Article  PubMed  CAS  Google Scholar 

  • Staley JP, Guthrie C (1998) Mechanical devices of the spliceosome: motors, clocks, springs, and things. Cell 92:315–326

    Article  PubMed  CAS  Google Scholar 

  • Stein L, Sternberg P, Durbin R, Thierry-Mieg J, Spieth J (2001) WormBase: network access to the genome and biology of Caenorhabditis elegans. Nucleic Acids Res 29:82–86

    Article  PubMed  CAS  Google Scholar 

  • Strom AC, Weis K (2001) Importin-beta-like nuclear transport receptors. Genome Biol 2:REVIEWS3008

    Google Scholar 

  • Toby GG, Golemis EA (2001) Using the yeast interaction trap and other two-hybrid-based approaches to study protein-protein interactions. Methods 24:201–217

    Article  PubMed  CAS  Google Scholar 

  • Tsuboi D, Qadota H, Kasuya K, Amano M, Kaibuchi K (2002) Isolation of the interacting molecules with GEX-3 by a novel functional screening. Biochem Biophys Res Commun 292:697–701

    Article  PubMed  CAS  Google Scholar 

  • Uetz P (2001) Two-hybrid arrays. Curr Opin Chem Biol 6:57–62

    Article  Google Scholar 

  • von Zelewsky T, Palladino F, Brunschwig K, Tobler H, Hajnal A, Muller F (2000) The C. elegans Mi-2 chromatin-remodelling proteins function in vulval cell fate determination. Development 127:5277–5284

    Google Scholar 

  • Walhout AJ, Boulton SJ, Vidal M (2000) Yeast two-hybrid systems and protein interaction mapping projects for yeast and worm. Yeast 17:88–94

    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 

  • Zhang Y, LeRoy G, Seelig HP, Lane WS, Reinberg D (1998) The dermatomyositis-specific autoantigen Mi2 is a component of a complex containing histone deacetylase and nucleosome remodeling activities. Cell 95:279–289

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y, Ng HH, Erdjument-Bromage H, Tempst P, Bird A, Reinberg D (1999) Analysis of the NuRD subunits reveals a histone deacetylase core complex and a connection with DNA methylation. Genes Dev 13:1924–35

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are grateful to Dr. Andrew Fire for providing GFP vectors, to Dr. Iain Johnstone for providing staged cDNA samples, to Dr. Robert Barstead for providing the cDNA library and to the Sequencing Unit (Dept. Biochemistry, University of Oxford) for carrying out all sequencing work. We thank Professor Kay Davies, Dr. Stefanos Christodoulou, Dr. Behrooz Esmaeili and Dr. Nick Owen for helpful discussions during the course of this work. This work was funded by the Medical Research Council of the UK.

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  1. MRC Functional Genetics Unit, Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK

    Paula R. Towers & David B. Sattelle

  2. Centre for Haematology, Bart’s and The London, Queen Mary’s School of Medicine, Institute of Cell and Molecular Science, 4 Newark Street, London, E1 2AT, UK

    Emma C. Burt

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  1. Emma C. Burt
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Correspondence to David B. Sattelle.

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Burt, E.C., Towers, P.R. & Sattelle, D.B. Caenorhabditis elegans in the study of SMN-interacting proteins: a role for SMI-1, an orthologue of human Gemin2 and the identification of novel components of the SMN complex. Invert Neurosci 6, 145–159 (2006). https://doi.org/10.1007/s10158-006-0027-x

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