CoChat
Sign Up
Cited by 221Neurogenetic and Muscular Disorders ResearchRNA Research and SplicingAmyotrophic Lateral Sclerosis Research

The Survival of Motor Neuron (SMN) Protein Interacts with the mRNA-Binding Protein HuD and Regulates Localization of Poly(A) mRNA in Primary Motor Neuron Axons

Open access
Fallini, Claudia, Zhang, Honglai, Su, Yuehang, Silani, Vincenzo, Singer, Robert H., Rossoll, Wilfried, Bassell, Gary J.·2011·The Journal of Neuroscience
DOI 10.1523/jneurosci.3631-10.2011PMC PMC3070748
Open this paper in CoChat — free accountOpen PDF

Abstract

Spinal muscular atrophy (SMA) results from reduced levels of the survival of motor neuron (SMN) protein, which has a well characterized function in spliceosomal small nuclear ribonucleoprotein assembly. Currently, it is not understood how deficiency of a housekeeping protein leads to the selective degeneration of spinal cord motor neurons. Numerous studies have shown that SMN is present in neuronal processes and has many interaction partners, including mRNA-binding proteins, suggesting a potential noncanonical role in axonal mRNA metabolism. In this study, we have established a novel technological approach using bimolecular fluorescence complementation (BiFC) and quantitative image analysis to characterize SMN-protein interactions in primary motor neurons. Consistent with biochemical studies on the SMN complex, BiFC analysis revealed that SMN dimerizes and interacts with Gemin2 in nuclear gems and axonal granules. In addition, using pull down assays, immunofluorescence, cell transfection, and BiFC, we characterized a novel interaction between SMN and the neuronal mRNA-binding protein HuD, which was dependent on the Tudor domain of SMN. A missense mutation in the SMN Tudor domain, which is known to cause SMA, impaired the interaction with HuD, but did not affect SMN axonal localization or self-association. Furthermore, time-lapse microscopy revealed SMN cotransport with HuD in live motor neurons. Importantly, SMN knockdown in primary motor neurons resulted in a specific reduction of both HuD protein and poly(A) mRNA levels in the axonal compartment. These findings reveal a noncanonical role for SMN whereby its interaction with mRNA-binding proteins may facilitate the localization of associated poly(A) mRNAs into axons.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1523/jneurosci.3631-10.2011

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The citation lists haven’t been fetched yet

References 73 · Cited by 221

Research works printed but not linked: 69

  1. Akamatsu W Fujihara H Mitsuhashi T Yano M Shibata S Hayakawa Y Okano HJ Sakakibara S Takano H Takano T Takahashi T Noda T Okano H The RNA-binding protein HuD regulates neuronal cell identity and maturation Proc Natl Acad Sci U S A 2005 102 4625 4630 15764704 PMC555491
  2. Anthony K Gallo JM Aberrant RNA processing events in neurological disorders Brain Res 2010 1338 67 77 20226177
  3. Aronov S Aranda G Behar L Ginzburg I Axonal tau mRNA localization coincides with tau protein in living neuronal cells and depends on axonal targeting signal J Neurosci 2001 21 6577 6587 11517247 PMC6763080
  4. Aronov S Aranda G Behar L Ginzburg I Visualization of translated tau protein in the axons of neuronal P19 cells and characterization of tau RNP granules J Cell Sci 2002 115 3817 3827 12235292
  5. Atlas R Behar L Elliott E Ginzburg I The insulin-like growth factor mRNA binding-protein IMP-1 and the Ras-regulatory protein G3BP associate with tau mRNA and HuD protein in differentiated P19 neuronal cells J Neurochem 2004 89 613 626 15086518
  6. Bassell GJ Powers CM Taneja KL Singer RH Single mRNAs Visualized by Ultrastructural In Situ Hybridization Are Principally Localized at Actin Filament Intersections in Fibroblasts Journal of Cell Biology 1994 126 863 876 7914201 PMC2120111
  7. Bassell GJ Zhang HL Byrd AL Femino AM Singer RH Taneja KL Lifshitz LM Herman IM Koisk KS Sorting of beta actin mRNA and protein to neurites and growth cones in culture Journal of Neuroscience 1998 18 251 265 9412505 PMC6793411
  8. Battle DJ Kasim M Yong J Lotti F Lau CK Mouaikel J Zhang Z Han K Wan L Dreyfuss G The SMN complex: an assembly machine for RNPs Cold Spring Harb Symp Quant Biol 2006 71 313 320 17381311
  9. Bolognani F Contente-Cuomo T Perrone-Bizzozero NI Novel recognition motifs and biological functions of the RNA-binding protein HuD revealed by genome-wide identification of its targets Nucleic Acids Res 2010 38 117 130 19846595 PMC2800223
  10. Bolognani F Merhege MA Twiss J Perrone-Bizzozero NI Dendritic localization of the RNA-binding protein HuD in hippocampal neurons: association with polysomes and upregulation during contextual learning Neurosci Lett 2004 371 152 157 15519747
  11. Bowerman M Shafey D Kothary R Smn depletion alters profilin II expression and leads to upregulation of the RhoA/ROCK pathway and defects in neuronal integrity J Mol Neurosci 2007 32 120 131 17873296
  12. Bowerman M Anderson CL Beauvais A Boyl PP Witke W Kothary R SMN, profilin IIa and plastin 3: a link between the deregulation of actin dynamics and SMA pathogenesis Mol Cell Neurosci 2009 42 66 74 19497369
  13. Briese M Esmaeili B Sattelle DB Is spinal muscular atrophy the result of defects in motor neuron processes? Bioessays 2005 27 946 957 16108074
  14. Buhler D Raker V Luhrmann R Fischer U Essential role for the tudor domain of SMN in spliceosomal U snRNP assembly: implications for spinal muscular atrophy Hum Mol Genet 1999 8 2351 2357 10556282
  15. Burghes AH Beattie CE Spinal muscular atrophy: why do low levels of survival motor neuron protein make motor neurons sick? Nat Rev Neurosci 2009 10 597 609 19584893 PMC2853768
  16. Burnett BG Munoz E Tandon A Kwon DY Sumner CJ Fischbeck KH Regulation of SMN protein stability Mol Cell Biol 2009 29 1107 1115 19103745 PMC2643817
  17. Chari A Paknia E Fischer U The role of RNP biogenesis in spinal muscular atrophy Curr Opin Cell Biol 2009 19286363
  18. Chung S Eckrich M Perrone-Bizzozero N Kohn DT Furneaux H The Elav-like proteins bind to a conserved regulatory element in the 3'-untranslated region of GAP-43 mRNA J Biol Chem 1997 272 6593 6598 9045688
  19. Costes SV Daelemans D Cho EH Dobbin Z Pavlakis G Lockett S Automatic and quantitative measurement of protein-protein colocalization in live cells Biophys J 2004 86 3993 4003 15189895 PMC1304300
  20. Cote J Richard S Tudor domains bind symmetrical dimethylated arginines J Biol Chem 2005 280 28476 28483 15955813
  21. Datta PK Chytil A Gorska AE Moses HL Identification of STRAP, a novel WD domain protein in transforming growth factor-beta signaling J Biol Chem 1998 273 34671 34674 9856985
  22. Denny JB Molecular Mechanisms, Biological Actions, and Neuropharmacology of the Growth-Associated Protein GAP-43 Curr Neuropharmacol 2006 4 293 304 18654638 PMC2475799
  23. Deschenes-Furry J Angus LM Belanger G Mwanjewe J Jasmin BJ Role of ELAV-like RNA-binding proteins HuD and HuR in the post-transcriptional regulation of acetylcholinesterase in neurons and skeletal muscle cells Chem Biol Interact 2005 157–158 43 49 16242680
  24. di Penta A Mercaldo V Florenzano F Munck S Ciotti MT Zalfa F Mercanti D Molinari M Bagni C Achsel T Dendritic LSm1/CBP80-mRNPs mark the early steps of transport commitment and translational control J Cell Biol 2009 184 423 435 19188494 PMC2646547
  25. Fallini C Bassell GJ Rossoll W High-efficiency transfection of cultured primary motor neurons to study protein localization, trafficking, and function Mol Neurodegener 2010 5 17 20406490 PMC2867961
  26. Fujiwara T Mori Y Chu DL Koyama Y Miyata S Tanaka H Yachi K Kubo T Yoshikawa H Tohyama M CARM1 regulates proliferation of PC12 cells by methylating HuD Mol Cell Biol 2006 26 2273 2285 16508003 PMC1430293
  27. Glinka M Herrmann T Funk N Havlicek S Rossoll W Winkler C Sendtner M The heterogeneous nuclear ribonucleoprotein-R is necessary for axonal beta-actin mRNA translocation in spinal motor neurons Hum Mol Genet 2010 19 1951 1966 20167579
  28. Grimmler M Otter S Peter C Muller F Chari A Fischer U Unrip, a factor implicated in cap-independent translation, associates with the cytosolic SMN complex and influences its intracellular localization Hum Mol Genet 2005 14 3099 3111 16159890
  29. Hahnen E Schonling J Rudnik-Schoneborn S Raschke H Zerres K Wirth B Missense mutations in exon 6 of the survival motor neuron gene in patients with spinal muscular atrophy (SMA) Hum Mol Genet 1997 6 821 825 9158159
  30. Helmken C Hofmann Y Schoenen F Oprea G Raschke H Rudnik-Schoneborn S Zerres K Wirth B Evidence for a modifying pathway in SMA discordant families: reduced SMN level decreases the amount of its interacting partners and Htra2-beta1 Hum Genet 2003 114 11 21 14520560
  31. Holt CE Bullock SL Subcellular mRNA localization in animal cells and why it matters Science 2009 326 1212 1216 19965463 PMC3785123
  32. Hu CD Chinenov Y Kerppola TK Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation Mol Cell 2002 9 789 798 11983170
  33. Jablonka S Bandilla M Wiese S Buhler D Wirth B Sendtner M Fischer U Co-regulation of survival of motor neuron (SMN) protein and its interactor SIP1 during development and in spinal muscular atrophy Hum Mol Genet 2001 10 497 505 11181573
  34. Kerppola TK Bimolecular fluorescence complementation: visualization of molecular interactions in living cells Methods Cell Biol 2008 85 431 470 18155474 PMC2829325
  35. Kerppola TK Visualization of molecular interactions using bimolecular fluorescence complementation analysis: characteristics of protein fragment complementation Chem Soc Rev 2009 38 2876 2886 19771334 PMC2980501
  36. Kolb SJ Battle DJ Dreyfuss G Molecular functions of the SMN complex J Child Neurol 2007 22 990 994 17761654
  37. Lemmens R Moore MJ Al-Chalabi A Brown RH Jr Robberecht W RNA metabolism and the pathogenesis of motor neuron diseases Trends Neurosci 2010 33 249 258 20227117
  38. Liu Q Fischer U Wang F Dreyfuss G The spinal muscular atrophy disease gene product, SMN, and its associated protein SIP1 are in a complex with spliceosomal snRNP proteins Cell 1997 90 1013 1021 9323129
  39. Lorson CL Strasswimmer J Yao JM Baleja JD Hahnen E Wirth B Le T Burghes AH Androphy EJ SMN oligomerization defect correlates with spinal muscular atrophy severity Nat Genet 1998 19 63 66 9590291
  40. Meister G Eggert C Fischer U SMN-mediated assembly of RNPs: a complex story Trends Cell Biol 2002 12 472 478 12441251
  41. Mobarak CD Anderson KD Morin M Beckel-Mitchener A Rogers SL Furneaux H King P Perrone-Bizzozero NI The RNA-binding protein HuD is required for GAP-43 mRNA stability, GAP-43 gene expression, and PKC-dependent neurite outgrowth in PC12 cells Mol Biol Cell 2000 11 3191 3203 10982410 PMC14985
  42. Monani UR Spinal muscular atrophy: a deficiency in a ubiquitous protein; a motor neuron-specific disease Neuron 2005 48 885 895 16364894
  43. Narayanan U Achsel T Luhrmann R Matera AG Coupled in vitro import of U snRNPs and SMN, the spinal muscular atrophy protein Mol Cell 2004 16 223 234 15494309
  44. Oprea GE Krober S McWhorter ML Rossoll W Muller S Krawczak M Bassell GJ Beattie CE Wirth B Plastin 3 is a protective modifier of autosomal recessive spinal muscular atrophy Science 2008 320 524 527 18440926 PMC4908855
  45. Otter S Grimmler M Neuenkirchen N Chari A Sickmann A Fischer U A comprehensive interaction map of the human survival of motor neuron (SMN) complex J Biol Chem 2007 282 5825 5833 17178713
  46. Pascale A Amadio M Quattrone A Defining a neuron: neuronal ELAV proteins Cell Mol Life Sci 2008 65 128 140 17928954 PMC11131659
  47. Perrone-Bizzozero N Bolognani F Role of HuD and other RNA-binding proteins in neural development and plasticity J Neurosci Res 2002 68 121 126 11948657
  48. Ramirez OA Vidal RL Tello JA Vargas KJ Kindler S Hartel S Couve A Dendritic assembly of heteromeric gamma-aminobutyric acid type B receptor subunits in hippocampal neurons J Biol Chem 2009 284 13077 13085 19276079 PMC2676040
  49. Ratti A Fallini C Colombrita C Pascale A Laforenza U Quattrone A Silani V Post-transcriptional regulation of neuro-oncological ventral antigen 1 by the neuronal RNA-binding proteins ELAV J Biol Chem 2008 283 7531 7541 18218628
  50. Robinow S Campos AR Yao KM White K The elav gene product of Drosophila, required in neurons, has three RNP consensus motifs Science 1988 242 1570 1572 3144044
  51. Rossoll W Bassell GJ Spinal Muscular Atrophy and a Model for Survival of Motor Neuron Protein Function in Axonal Ribonucleoprotein Complexes Results Probl Cell Differ 2009 PMC3718852 19343312
  52. Rossoll W Kroning AK Ohndorf UM Steegborn C Jablonka S Sendtner M Specific interaction of Smn, the spinal muscular atrophy determining gene product, with hnRNP-R and gry-rbp/hnRNP-Q: a role for Smn in RNA processing in motor axons? Hum Mol Genet 2002 11 93 105 11773003
  53. Rossoll W Jablonka S Andreassi C Kroning AK Karle K Monani UR Sendtner M Smn, the spinal muscular atrophy-determining gene product, modulates axon growth and localization of beta-actin mRNA in growth cones of motoneurons J Cell Biol 2003 163 801 812 14623865 PMC2173668
  54. Selenko P Sprangers R Stier G Buhler D Fischer U Sattler M SMN tudor domain structure and its interaction with the Sm proteins Nat Struct Biol 2001 8 27 31 11135666
  55. Sendtner M Molecular mechanisms in spinal muscular atrophy: models and perspectives Curr Opin Neurol 2001 14 629 634 11562575
  56. Shyu YJ Liu H Deng X Hu CD Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions Biotechniques 2006 40 61 66 16454041
  57. Sleeman JE Lamond AI Newly assembled snRNPs associate with coiled bodies before speckles, suggesting a nuclear snRNP maturation pathway Curr Biol 1999 9 1065 1074 10531003
  58. Smith CL Afroz R Bassell GJ Furneaux HM Perrone-Bizzozero NI Burry RW GAP-43 mRNA in growth cones is associated with HuD and ribosomes J Neurobiol 2004 61 222 235 15389607
  59. Tadesse H Deschenes-Furry J Boisvenue S Cote J KH-type splicing regulatory protein interacts with survival motor neuron protein and is misregulated in spinal muscular atrophy Hum Mol Genet 2008 17 506 524 17998247
  60. Tiruchinapalli DM Ehlers MD Keene JD Activity-dependent expression of RNA binding protein HuD and its association with mRNAs in neurons RNA Biol 2008 5 157 168 18769135
  61. Tretyakova I Zolotukhin AS Tan W Bear J Propst F Ruthel G Felber BK Nuclear export factor family protein participates in cytoplasmic mRNA trafficking J Biol Chem 2005 280 31981 31990 16014633
  62. Vidi PA Chemel BR Hu CD Watts VJ Ligand-dependent oligomerization of dopamine D(2) and adenosine A(2A) receptors in living neuronal cells Mol Pharmacol 2008 74 544 551 18524886
  63. Wang J Dreyfuss G A cell system with targeted disruption of the SMN gene: functional conservation of the SMN protein and dependence of Gemin2 on SMN J Biol Chem 2001 276 9599 9605 11121410
  64. Yuan Z Gong S Luo J Zheng Z Song B Ma S Guo J Hu C Thiel G Vinson C Hu CD Wang Y Li M Opposing roles for ATF2 and c-Fos in c-Jun-mediated neuronal apoptosis Mol Cell Biol 2009 29 2431 2442 19255142 PMC2668374
  65. Zacharias DA Violin JD Newton AC Tsien RY Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells Science 2002 296 913 916 11988576
  66. Zhang H Xing L Rossoll W Wichterle H Singer RH Bassell GJ Multiprotein complexes of the survival of motor neuron protein SMN with Gemins traffic to neuronal processes and growth cones of motor neurons J Neurosci 2006 26 8622 8632 16914688 PMC4956918
  67. Zhang HL Singer RH Bassell GJ Neurotrophin regulation of beta-actin mRNA and protein localization within growth cones J Cell Biol 1999 147 59 70 10508855 PMC2164987
  68. Zhang HL Pan F Hong D Shenoy SM Singer RH Bassell GJ Active transport of the survival motor neuron protein and the role of exon-7 in cytoplasmic localization J Neurosci 2003 23 6627 6637 12878704 PMC6740639
  69. Zhang Z Lotti F Dittmar K Younis I Wan L Kasim M Dreyfuss G SMN deficiency causes tissue-specific perturbations in the repertoire of snRNAs and widespread defects in splicing Cell 2008 133 585 600 18485868 PMC2446403

Authors
Fallini, C., Zhang, H., Su, Y., Silani, V., Singer, R. H., Rossoll, W., & Bassell, G. J.
Affiliations
  • Emory University (US)
  • Albert Einstein College of Medicine (US)
  • University of Milan (IT)
Corresponding author
  • Rossoll, Wilfried
  • Bassell, Gary J.
ORCID
Venue
The Journal of Neuroscience
Publisher
Society for Neuroscience
Published
2011-03-09
Volume
31
Issue
10
Pages
3914-3925
ISSN
  • 0270-6474 (ISSN-L)
  • 1529-2401
DOI
10.1523/jneurosci.3631-10.2011
PMC
PMC3070748
OpenAlex
W2056116259
Cited by
221
References
73
Citations by year
2026: 10 · 2025: 3 · 2024: 7 · 2023: 14 · 2022: 14 · 2021: 13 · 2020: 21 · 2019: 7 · 2018: 14 · 2017: 23 · 2016: 19 · 2015: 8 · 2014: 18 · 2013: 24 · 2012: 18
FWCI
11.04
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Journal of Neuroscience Nursing
Funders
  • Spinal Muscular Atrophy Foundation
  • Emory University (P30NS055077)
  • National Institutes of Health (NS066030)
  • National Institute of General Medical Sciences (R01GM084364)
  • National Institute of Neurological Disorders and Stroke (R21NS066030, P30NS055077, NS066030)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (R01HD055835, HD055835)
Topics
  • Health Sciences › Medicine › Genetics › Neurogenetic and Muscular Disorders Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Health Sciences › Medicine › Neurology › Amyotrophic Lateral Sclerosis Research
Keywords
survival motor neuron protein, HUD, spinal muscular atrophy, poly(A)+ mRNA, mRNA-binding proteins, Tudor domain, Gemin2, motor neurons, axonal transport
MeSH terms
ELAV-Like Protein 4, Animals, Axons, Chick Embryo, Fluorescent Antibody Technique, Hippocampus, Humans, Motor Neurons, Poly A, Blotting, Western, Prosencephalon, In Situ Hybridization, Fluorescence, Poly(A)-Binding Proteins, Immunoprecipitation, Mice, Rats, ELAV Proteins, SMN Complex Proteins, HEK293 Cells

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review