Acta Neuropathologica

, Volume 124, Issue 5, pp 705–716 | Cite as

Transportin 1 accumulates specifically with FET proteins but no other transportin cargos in FTLD-FUS and is absent in FUS inclusions in ALS with FUS mutations

  • Manuela Neumann
  • Chiara F. Valori
  • Olaf Ansorge
  • Hans A. Kretzschmar
  • David G. Munoz
  • Hirofumi Kusaka
  • Osamu Yokota
  • Kenji Ishihara
  • Lee-Cyn Ang
  • Juan M. Bilbao
  • Ian R. A. Mackenzie
Original Paper

Abstract

Accumulation of the DNA/RNA binding protein fused in sarcoma (FUS) as inclusions in neurons and glia is the pathological hallmark of amyotrophic lateral sclerosis patients with mutations in FUS (ALS-FUS) as well as in several subtypes of frontotemporal lobar degeneration (FTLD-FUS), which are not associated with FUS mutations. Despite some overlap in the phenotype and neuropathology of FTLD-FUS and ALS-FUS, significant differences of potential pathomechanistic relevance were recently identified in the protein composition of inclusions in these conditions. While ALS-FUS showed only accumulation of FUS, inclusions in FTLD-FUS revealed co-accumulation of all members of the FET protein family, that include FUS, Ewing’s sarcoma (EWS) and TATA-binding protein-associated factor 15 (TAF15) suggesting a more complex disturbance of transportin-mediated nuclear import of proteins in FTLD-FUS compared to ALS-FUS. To gain more insight into the mechanisms of inclusion body formation, we investigated the role of Transportin 1 (Trn1) as well as 13 additional cargo proteins of Transportin in the spectrum of FUS-opathies by immunohistochemistry and biochemically. FUS-positive inclusions in six ALS-FUS cases including four different mutations did not label for Trn1. In sharp contrast, the FET-positive pathology in all FTLD-FUS subtypes was also strongly labeled for Trn1 and often associated with a reduction in the normal nuclear staining of Trn1 in inclusion bearing cells, while no biochemical changes of Trn1 were detectable in FTLD-FUS. Notably, despite the dramatic changes in the subcellular distribution of Trn1 in FTLD-FUS, alterations of its cargo proteins were restricted to FET proteins and no changes in the normal physiological staining of 13 additional Trn1 targets, such as hnRNPA1, PAPBN1 and Sam68, were observed in FTLD-FUS. These data imply a specific dysfunction in the interaction between Trn1 and FET proteins in the inclusion body formation in FTLD-FUS. Moreover, the absence of Trn1 in ALS-FUS provides further evidence that ALS-FUS and FTLD-FUS have different underlying pathomechanisms.

Keywords

Transportin FUS TAF15 EWS Amyotrophic lateral sclerosis Frontotemporal dementia 

Notes

Acknowledgments

We thank Margaret Luk and Jay Tracy for their excellent technical assistance. This work was supported by grants from the Swiss National Science Foundation (31003A-132864, MN); the Synapsis Foundation (MN); the Canadian Institutes of Health Research (74580 and 179009, IM), the Pacific Alzheimer’s Research Foundation (C06-01, IM); and the NIHR Oxford Biomedical Research Centre (OA).

Supplementary material

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Supplementary material 1 (PDF 1101 kb)

References

  1. 1.
    Belyanskaya LL, Gehrig PM, Gehring H (2001) Exposure on cell surface and extensive arginine methylation of Ewing sarcoma (EWS) protein. J Biol Chem 276:18681–18687PubMedCrossRefGoogle Scholar
  2. 2.
    Belyanskaya LL, Delattre O, Gehring H (2003) Expression and subcellular localization of Ewing sarcoma (EWS) protein is affected by the methylation process. Exp Cell Res 288:374–381PubMedCrossRefGoogle Scholar
  3. 3.
    Blair IP, Williams KL, Warraich ST et al (2010) FUS mutations in amyotrophic lateral sclerosis: clinical, pathological, neurophysiological and genetic analysis. J Neurol Neurosurg Psychiatry 81:639–645PubMedCrossRefGoogle Scholar
  4. 4.
    Brelstaff J, Lashley T, Holton JL et al (2011) Transportin1: a marker of FTLD-FUS. Acta Neuropathol 122:591–600PubMedCrossRefGoogle Scholar
  5. 5.
    Davidson YS, Robinson AC, Hu Q et al (2012) Nuclear carrier and RNA binding proteins in frontotemporal lobar degeneration associated with fused in sarcoma (FUS) pathological changes. Neuropathol Appl Neurobiol. doi:10.1111/j.1365-2990.2012.01274.x
  6. 6.
    Doi H, Koyano S, Suzuki Y, Nukina N, Kuroiwa Y (2010) The RNA-binding protein FUS/TLS is a common aggregate-interacting protein in polyglutamine diseases. Neurosci Res 66:131–133PubMedCrossRefGoogle Scholar
  7. 7.
    Dormann D, Rodde R, Edbauer D et al (2010) ALS-associated fused in sarcoma (FUS) mutations disrupt Transportin-mediated nuclear import. EMBO J 29:2841–2857PubMedCrossRefGoogle Scholar
  8. 8.
    Fronz K, Guttinger S, Burkert K et al (2011) Arginine methylation of the nuclear poly(a) binding protein weakens the interaction with its nuclear import receptor, transportin. J Biol Chem 286:32986–32994PubMedCrossRefGoogle Scholar
  9. 9.
    Groen EJ, van Es MA, van Vught PW et al (2010) FUS mutations in familial amyotrophic lateral sclerosis in the Netherlands. Arch Neurol 67:224–230PubMedCrossRefGoogle Scholar
  10. 10.
    Hewitt C, Kirby J, Highley JR et al (2010) Novel FUS/TLS mutations and pathology in familial and sporadic amyotrophic lateral sclerosis. Arch Neurol 67:455–461PubMedCrossRefGoogle Scholar
  11. 11.
    Ito D, Seki M, Tsunoda Y, Uchiyama H, Suzuki N (2010) Nuclear transport impairment of amyotrophic lateral sclerosis-linked mutations in FUS/TLS. Ann Neurol 69(1):152–162PubMedCrossRefGoogle Scholar
  12. 12.
    Jobert L, Argentini M, Tora L (2009) PRMT1 mediated methylation of TAF15 is required for its positive gene regulatory function. Exp Cell Res 315:1273–1286PubMedCrossRefGoogle Scholar
  13. 13.
    Kino Y, Washizu C, Aquilanti E et al (2011) Intracellular localization and splicing regulation of FUS/TLS are variably affected by amyotrophic lateral sclerosis-linked mutations. Nucleic Acids Res 39:2781–2798PubMedCrossRefGoogle Scholar
  14. 14.
    Kovar H (2011) Dr. Jekyll and Mr. Hyde: the two faces of the FUS/EWS/TAF15 Protein Family. Sarcoma 837474. doi:10.1155/2011/837474
  15. 15.
    Kwiatkowski TJ Jr, Bosco DA, Leclerc AL et al (2009) Mutations in the FUS/TLS gene on chromosome 16 cause familial amyotrophic lateral sclerosis. Science 323:1205–1208PubMedCrossRefGoogle Scholar
  16. 16.
    Lashley T, Rohrer JD, Bandopadhyay R et al (2011) A comparative clinical, pathological, biochemical and genetic study of fused in sarcoma proteinopathies. Brain 134:2548–2564PubMedCrossRefGoogle Scholar
  17. 17.
    Law WJ, Cann KL, Hicks GG (2006) TLS, EWS and TAF15: a model for transcriptional integration of gene expression. Brief Funct Genomic Proteomic 5:8–14PubMedCrossRefGoogle Scholar
  18. 18.
    Lee BJ, Cansizoglu AE, Suel KE et al (2006) Rules for nuclear localization sequence recognition by karyopherin beta 2. Cell 126:543–558PubMedCrossRefGoogle Scholar
  19. 19.
    Mackenzie IR, Neumann M, Bigio EH et al (2010) Nomenclature and nosology for neuropathologic subtypes of frontotemporal lobar degeneration: an update. Acta Neuropathol 119:1–4PubMedCrossRefGoogle Scholar
  20. 20.
    Mackenzie IR, Rademakers R, Neumann M (2010) TDP-43 and FUS in amyotrophic lateral sclerosis and frontotemporal dementia. Lancet Neurol 9:995–1007PubMedCrossRefGoogle Scholar
  21. 21.
    Mackenzie IR, Munoz DG, Kusaka H et al (2011) Distinct pathological subtypes of FTLD-FUS. Acta Neuropathol 121:207–218PubMedCrossRefGoogle Scholar
  22. 22.
    Mackenzie IR, Neumann M, Baborie A et al (2011) A harmonized classification system for FTLD-TDP pathology. Acta Neuropathol 122:111–113PubMedCrossRefGoogle Scholar
  23. 23.
    Mackenzie IRA, Ansorge O, Strong M et al (2011) Pathological heterogeneity in amyotrophic lateral sclerosis with FUS mutations: two distinct patterns correlating with disease severity and mutation. Acta Neuropathol 122:87–98PubMedCrossRefGoogle Scholar
  24. 24.
    Munoz DG, Neumann M, Kusaka H et al (2009) FUS pathology in basophilic inclusion body disease. Acta Neuropathol 118:617–627PubMedCrossRefGoogle Scholar
  25. 25.
    Neumann M, Rademakers R, Roeber S et al (2009) A new subtype of frontotemporal lobar degeneration with FUS pathology. Brain 132:2922–2931PubMedCrossRefGoogle Scholar
  26. 26.
    Neumann M, Roeber S, Kretzschmar HA et al (2009) Abundant FUS-immunoreactive pathology in neuronal intermediate filament inclusion disease. Acta Neuropathol 118:605–616PubMedCrossRefGoogle Scholar
  27. 27.
    Neumann M, Bentmann E, Dormann D et al (2011) FET proteins TAF15 and EWS are selective markers that distinguish FTLD with FUS pathology from amyotrophic lateral sclerosis with FUS mutations. Brain 134:2595–2609PubMedCrossRefGoogle Scholar
  28. 28.
    Page T, Gitcho MA, Mosaheb S et al (2011) FUS immunogold labeling TEM analysis of the neuronal cytoplasmic inclusions of neuronal intermediate filament inclusion disease: a frontotemporal lobar degeneration with FUS proteinopathy. J Mol Neurosci 45:409–421PubMedCrossRefGoogle Scholar
  29. 29.
    Rademakers R, Stewart H, DeJesus-Hernandez M et al (2010) FUS gene mutations in familial and sporadic amyotrophic lateral sclerosis. Muscle Nerve 42:170–176PubMedCrossRefGoogle Scholar
  30. 30.
    Rappsilber J, Friesen WJ, Paushkin S, Dreyfuss G, Mann M (2003) Detection of arginine dimethylated peptides by parallel precursor ion scanning mass spectrometry in positive ion mode. Anal Chem 75:3107–3114PubMedCrossRefGoogle Scholar
  31. 31.
    Rohrer JD, Lashley T, Holton J et al (2011) The clinical and neuroanatomical phenotype of FUS associated frontotemporal lobar degeneration. J Neurol Neurosurg Psychiatry 82:1405–1407PubMedCrossRefGoogle Scholar
  32. 32.
    Snowden JS, Hu Q, Rollinson S et al (2011) The most common type of FTLD-FUS (aFTLD-U) is associated with a distinct clinical form of frontotemporal dementia but is not related to mutations in the FUS gene. Acta Neuropathol 122:99–110PubMedCrossRefGoogle Scholar
  33. 33.
    Suel KE, Gu H, Chook YM (2008) Modular organization and combinatorial energetics of proline-tyrosine nuclear localization signals. PLoS Biol 6:e137PubMedCrossRefGoogle Scholar
  34. 34.
    Tan AY, Manley JL (2009) The TET family of proteins: functions and roles in disease. J Mol Cell Biol 1:82–92PubMedCrossRefGoogle Scholar
  35. 35.
    Tradewell ML, Yu Z, Tibshirani M et al (2012) Arginine methylation by PRMT1 regulates nuclear-cytoplasmic localization and toxicity of FUS/TLS harbouring ALS-linked mutations. Hum Mol Genet 21:136–149PubMedCrossRefGoogle Scholar
  36. 36.
    Urwin H, Josephs KA, Rohrer JD et al (2010) FUS pathology defines the majority of tau- and TDP-43-negative frontotemporal lobar degeneration. Acta Neuropathol 120:33–41PubMedCrossRefGoogle Scholar
  37. 37.
    Vance C, Rogelj B, Hortobagyi T et al (2009) Mutations in FUS, an RNA processing protein, cause familial amyotrophic lateral sclerosis type 6. Science 323:1208–1211PubMedCrossRefGoogle Scholar
  38. 38.
    Woulfe J, Gray DA, Mackenzie IR (2010) FUS-immunoreactive intranuclear inclusions in neurodegenerative disease. Brain Pathol 20:589–597PubMedCrossRefGoogle Scholar
  39. 39.
    Zakaryan RP, Gehring H (2006) Identification and characterization of the nuclear localization/retention signal in the EWS proto-oncoprotein. J Mol Biol 363:27–38PubMedCrossRefGoogle Scholar
  40. 40.
    Zinszner H, Sok J, Immanuel D, Yin Y, Ron D (1997) TLS (FUS) binds RNA in vivo and engages in nucleo-cytoplasmic shuttling. J Cell Sci 110:1741–1750PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Manuela Neumann
    • 1
    • 2
    • 3
  • Chiara F. Valori
    • 1
  • Olaf Ansorge
    • 4
  • Hans A. Kretzschmar
    • 5
  • David G. Munoz
    • 6
  • Hirofumi Kusaka
    • 7
  • Osamu Yokota
    • 8
  • Kenji Ishihara
    • 9
  • Lee-Cyn Ang
    • 10
  • Juan M. Bilbao
    • 11
  • Ian R. A. Mackenzie
    • 12
  1. 1.Institute of NeuropathologyUniversity Hospital ZurichZurichSwitzerland
  2. 2.Department of NeuropathologyUniversity of TübingenTübingenGermany
  3. 3.DZNE, German Center for Neurodegenerative DiseasesTübingenGermany
  4. 4.Department of NeuropathologyJohn Radcliffe HospitalOxfordUK
  5. 5.Center for Neuropathology and Prion ResearchLudwig-Maximilians-UniversityMunichGermany
  6. 6.Department of Laboratory Medicine and Pathobiology, Li Ka Shing Knowledge Institute, St. Michael’s HospitalUniversity of TorontoTorontoCanada
  7. 7.Department of NeurologyKansai Medical UniversityOsakaJapan
  8. 8.Department of NeuropsychiatryOkayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesOkayamaJapan
  9. 9.Department of NeurologyShowa University School of MedicineTokyoJapan
  10. 10.Department of PathologyLondon Health Sciences CentreLondonCanada
  11. 11.Department of PathologySunnybrook Health Sciences CentreTorontoCanada
  12. 12.Department of Pathology, Vancouver General HospitalUniversity of British ColumbiaVancouverCanada

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