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 (original) (raw)
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.
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References
- Belyanskaya LL, Gehrig PM, Gehring H (2001) Exposure on cell surface and extensive arginine methylation of Ewing sarcoma (EWS) protein. J Biol Chem 276:18681–18687
Article PubMed CAS Google Scholar - 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 288:374–381
Article PubMed CAS Google Scholar - 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 81:639–645
Article PubMed Google Scholar - Brelstaff J, Lashley T, Holton JL et al (2011) Transportin1: a marker of FTLD-FUS. Acta Neuropathol 122:591–600
Article PubMed CAS Google Scholar - 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
- 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 66:131–133
Article PubMed CAS Google Scholar - Dormann D, Rodde R, Edbauer D et al (2010) ALS-associated fused in sarcoma (FUS) mutations disrupt Transportin-mediated nuclear import. EMBO J 29:2841–2857
Article PubMed CAS Google Scholar - 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 286:32986–32994
Article PubMed CAS Google Scholar - Groen EJ, van Es MA, van Vught PW et al (2010) FUS mutations in familial amyotrophic lateral sclerosis in the Netherlands. Arch Neurol 67:224–230
Article PubMed Google Scholar - Hewitt C, Kirby J, Highley JR et al (2010) Novel FUS/TLS mutations and pathology in familial and sporadic amyotrophic lateral sclerosis. Arch Neurol 67:455–461
Article PubMed Google Scholar - 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):152–162
Article PubMed Google Scholar - Jobert L, Argentini M, Tora L (2009) PRMT1 mediated methylation of TAF15 is required for its positive gene regulatory function. Exp Cell Res 315:1273–1286
Article PubMed CAS Google Scholar - 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 39:2781–2798
Article PubMed CAS Google Scholar - 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
- 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 323:1205–1208
Article PubMed CAS Google Scholar - Lashley T, Rohrer JD, Bandopadhyay R et al (2011) A comparative clinical, pathological, biochemical and genetic study of fused in sarcoma proteinopathies. Brain 134:2548–2564
Article PubMed Google Scholar - Law WJ, Cann KL, Hicks GG (2006) TLS, EWS and TAF15: a model for transcriptional integration of gene expression. Brief Funct Genomic Proteomic 5:8–14
Article PubMed CAS Google Scholar - Lee BJ, Cansizoglu AE, Suel KE et al (2006) Rules for nuclear localization sequence recognition by karyopherin beta 2. Cell 126:543–558
Article PubMed CAS Google Scholar - Mackenzie IR, Neumann M, Bigio EH et al (2010) Nomenclature and nosology for neuropathologic subtypes of frontotemporal lobar degeneration: an update. Acta Neuropathol 119:1–4
Article PubMed Google Scholar - Mackenzie IR, Rademakers R, Neumann M (2010) TDP-43 and FUS in amyotrophic lateral sclerosis and frontotemporal dementia. Lancet Neurol 9:995–1007
Article PubMed CAS Google Scholar - Mackenzie IR, Munoz DG, Kusaka H et al (2011) Distinct pathological subtypes of FTLD-FUS. Acta Neuropathol 121:207–218
Article PubMed Google Scholar - Mackenzie IR, Neumann M, Baborie A et al (2011) A harmonized classification system for FTLD-TDP pathology. Acta Neuropathol 122:111–113
Article PubMed Google Scholar - 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 122:87–98
Article PubMed Google Scholar - Munoz DG, Neumann M, Kusaka H et al (2009) FUS pathology in basophilic inclusion body disease. Acta Neuropathol 118:617–627
Article PubMed CAS Google Scholar - Neumann M, Rademakers R, Roeber S et al (2009) A new subtype of frontotemporal lobar degeneration with FUS pathology. Brain 132:2922–2931
Article PubMed Google Scholar - Neumann M, Roeber S, Kretzschmar HA et al (2009) Abundant FUS-immunoreactive pathology in neuronal intermediate filament inclusion disease. Acta Neuropathol 118:605–616
Article PubMed CAS Google Scholar - 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 134:2595–2609
Article PubMed Google Scholar - 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 45:409–421
Article PubMed CAS Google Scholar - Rademakers R, Stewart H, DeJesus-Hernandez M et al (2010) FUS gene mutations in familial and sporadic amyotrophic lateral sclerosis. Muscle Nerve 42:170–176
Article PubMed CAS Google Scholar - 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 75:3107–3114
Article PubMed CAS Google Scholar - Rohrer JD, Lashley T, Holton J et al (2011) The clinical and neuroanatomical phenotype of FUS associated frontotemporal lobar degeneration. J Neurol Neurosurg Psychiatry 82:1405–1407
Article PubMed Google Scholar - 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 122:99–110
Article PubMed CAS Google Scholar - Suel KE, Gu H, Chook YM (2008) Modular organization and combinatorial energetics of proline-tyrosine nuclear localization signals. PLoS Biol 6:e137
Article PubMed Google Scholar - Tan AY, Manley JL (2009) The TET family of proteins: functions and roles in disease. J Mol Cell Biol 1:82–92
Article PubMed CAS Google Scholar - 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 21:136–149
Article PubMed Google Scholar - 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 120:33–41
Article PubMed Google Scholar - Vance C, Rogelj B, Hortobagyi T et al (2009) Mutations in FUS, an RNA processing protein, cause familial amyotrophic lateral sclerosis type 6. Science 323:1208–1211
Article PubMed CAS Google Scholar - Woulfe J, Gray DA, Mackenzie IR (2010) FUS-immunoreactive intranuclear inclusions in neurodegenerative disease. Brain Pathol 20:589–597
Article PubMed CAS Google Scholar - Zakaryan RP, Gehring H (2006) Identification and characterization of the nuclear localization/retention signal in the EWS proto-oncoprotein. J Mol Biol 363:27–38
Article PubMed CAS Google Scholar - 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 110:1741–1750
PubMed CAS Google Scholar
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).
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Authors and Affiliations
- Institute of Neuropathology, University Hospital Zurich, Zurich, Switzerland
Manuela Neumann & Chiara F. Valori - Department of Neuropathology, University of Tübingen, Calwerstr. 3, 72076, Tübingen, Germany
Manuela Neumann - DZNE, German Center for Neurodegenerative Diseases, Tübingen, Germany
Manuela Neumann - Department of Neuropathology, John Radcliffe Hospital, Oxford, UK
Olaf Ansorge - Center for Neuropathology and Prion Research, Ludwig-Maximilians-University, Munich, Germany
Hans A. Kretzschmar - Department of Laboratory Medicine and Pathobiology, Li Ka Shing Knowledge Institute, St. Michael’s Hospital, University of Toronto, Toronto, Canada
David G. Munoz - Department of Neurology, Kansai Medical University, Osaka, Japan
Hirofumi Kusaka - Department of Neuropsychiatry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan
Osamu Yokota - Department of Neurology, Showa University School of Medicine, Tokyo, Japan
Kenji Ishihara - Department of Pathology, London Health Sciences Centre, London, ON, Canada
Lee-Cyn Ang - Department of Pathology, Sunnybrook Health Sciences Centre, Toronto, ON, Canada
Juan M. Bilbao - Department of Pathology, Vancouver General Hospital, University of British Columbia, Vancouver, Canada
Ian R. A. Mackenzie
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Neumann, M., Valori, C.F., Ansorge, O. et al. 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.Acta Neuropathol 124, 705–716 (2012). https://doi.org/10.1007/s00401-012-1020-6
- Received: 22 May 2012
- Accepted: 16 July 2012
- Published: 28 July 2012
- Issue Date: November 2012
- DOI: https://doi.org/10.1007/s00401-012-1020-6