Skip to main content

A role for SUMOylation in the Formation and Cellular Localization of TDP-43 Aggregates in Amyotrophic Lateral Sclerosis

Molecular Neurobiology volume 57, pages 1361–1373 (2020)Cite this article

Abstract

In amyotrophic lateral sclerosis, motor neurons undergoing degeneration are characterized by the presence of cytoplasmic aggregates containing TDP-43 protein. SUMOylation, a posttranslational modification of proteins, has been previously implicated in the formation of aggregates positives for SOD1, another protein enriched in a subset of ALS patients. We show in this study that TDP-43 is also a target of SUMOylation. The inhibition of the first step of the SUMOylation process by anacardic acid significantly reduces the presence of TDP-43 aggregates and improves neuritogenesis and cell viability in vitro. Interestingly, the mutation of the unique SUMOylation site on TDP-43, using site-directed mutagenesis, modifies the intracellular localization of TDP-43 aggregates. Instead of being cytoplasmic where they are associated with toxic effects, they are located inside the nucleus. This change of localization results in improvement in cell viability and in global cellular functions. Our results implicate the SUMOylation site of TDP-43 in the formation of cytoplasmic TDP-43 aggregates, a hallmark of ALS, and thus identifies this region as a new target for novel therapeutic strategies.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price includes VAT (USA)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Abbreviations

ALS:

Amyotrophic lateral sclerosis

AA:

Anacardic acid

FTD:

Fronto-temporal dementia

PTM:

Posttranslational modifications

RBPs:

RNA-binding proteins

SUMO:

Small ubiquitin-like modifier

SOD1:

Superoxide dismutase 1

TARDBP:

TAR DNA-binding protein

MAPT:

Microtubule-associated protein Tau

GAP43:

Growth-associated protein

MAP 2:

Microtubule-associated protein 2

VAChT:

Vesicular acetylcholine transporter

ChAT:

Choline O-acetyltransferase

AChE:

Acetylcholinesterase

References

  1. Brown RH, Al-Chalabi A (2017) Amyotrophic lateral sclerosis. N Engl J Med 377:162–172. https://doi.org/10.1056/NEJMra1603471

    CAS  Article  PubMed  Google Scholar 

  2. Neumann M, Sampathu DM, Kwong LK et al (2006) Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science 314:130–133. https://doi.org/10.1126/science.1134108

    CAS  Article  PubMed  Google Scholar 

  3. Arai T, Hasegawa M, Akiyama H, Ikeda K, Nonaka T, Mori H, Mann D, Tsuchiya K et al (2006) TDP-43 is a component of ubiquitin-positive tau-negative inclusions in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Biochem Biophys Res Commun 351:602–611. https://doi.org/10.1016/j.bbrc.2006.10.093

    CAS  Article  PubMed  Google Scholar 

  4. Mackenzie IR, Rademakers R, Neumann M (2010) TDP-43 and FUS in amyotrophic lateral sclerosis and frontotemporal dementia. Lancet Neurol 9:995–1007. https://doi.org/10.1016/S1474-4422(10)70195-2

    CAS  Article  PubMed  Google Scholar 

  5. Millecamps S, Salachas F, Cazeneuve C, Gordon P, Bricka B, Camuzat A, Guillot-Noël L, Russaouen O et al (2010) SOD1, ANG, VAPB, TARDBP, and FUS mutations in familial amyotrophic lateral sclerosis: genotype-phenotype correlations. J Med Genet 47:554–560. https://doi.org/10.1136/jmg.2010.077180

    CAS  Article  PubMed  Google Scholar 

  6. Müller K, Brenner D, Weydt P et al (2018) Comprehensive analysis of the mutation spectrum in 301 German ALS families. J Neurol Neurosurg Psychiatry. https://doi.org/10.1136/jnnp-2017-317611

    Article  Google Scholar 

  7. Wegorzewska I, Bell S, Cairns NJ, Miller TM, Baloh RH (2009) TDP-43 mutant transgenic mice develop features of ALS and frontotemporal lobar degeneration. Proc Natl Acad Sci U S A 106:18809–18814. https://doi.org/10.1073/pnas.0908767106

    Article  PubMed  PubMed Central  Google Scholar 

  8. Xu Y-F, Gendron TF, Zhang Y-J, Lin WL, D'Alton S, Sheng H, Casey MC, Tong J et al (2010) Wild-type human TDP-43 expression causes TDP-43 phosphorylation, mitochondrial aggregation, motor deficits, and early mortality in transgenic mice. J Neurosci 30:10851–10859. https://doi.org/10.1523/JNEUROSCI.1630-10.2010

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. Scotter EL, Chen H-J, Shaw CE (2015) TDP-43 Proteinopathy and ALS: insights into disease mechanisms and therapeutic targets. Neurother J Am Soc Exp Neurother 12:352–363. https://doi.org/10.1007/s13311-015-0338-x

    CAS  Article  Google Scholar 

  10. Ederle H, Dormann D (2017) TDP-43 and FUS en route from the nucleus to the cytoplasm. FEBS Lett. https://doi.org/10.1002/1873-3468.12646

    CAS  Article  PubMed  Google Scholar 

  11. Svahn AJ, Don EK, Badrock AP, Cole NJ, Graeber MB, Yerbury JJ, Chung R, Morsch M (2018) Nucleo-cytoplasmic transport of TDP-43 studied in real time: impaired microglia function leads to axonal spreading of TDP-43 in degenerating motor neurons. Acta Neuropathol (Berl) 136:445–459. https://doi.org/10.1007/s00401-018-1875-2

    CAS  Article  Google Scholar 

  12. Birsa N, Bentham MP, Fratta P (2019) Cytoplasmic functions of TDP-43 and FUS and their role in ALS. Semin Cell Dev Biol. https://doi.org/10.1016/j.semcdb.2019.05.023

  13. Baloh RH (2011) TDP-43: the relationship between protein aggregation and neurodegeneration in amyotrophic lateral sclerosis and frontotemporal lobar degeneration: Protein autoregulation mechanisms. FEBS J 278:3539–3549. https://doi.org/10.1111/j.1742-4658.2011.08256.x

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. Blokhuis AM, Groen EJN, Koppers M, van den Berg L, Pasterkamp RJ (2013) Protein aggregation in amyotrophic lateral sclerosis. Acta Neuropathol (Berl) 125:777–794. https://doi.org/10.1007/s00401-013-1125-6

    CAS  Article  Google Scholar 

  15. Buratti E (2018) TDP-43 post-translational modifications in health and disease. Expert Opin Ther Targets 22:279–293. https://doi.org/10.1080/14728222.2018.1439923

    CAS  Article  PubMed  Google Scholar 

  16. Dangoumau A, Veyrat-Durebex C, Blasco H et al (2013) Protein SUMOylation, an emerging pathway in amyotrophic lateral sclerosis. Int J Neurosci 123:366–374. https://doi.org/10.3109/00207454.2012.761984

    CAS  Article  PubMed  Google Scholar 

  17. Dangoumau A, Marouillat S, Burlaud Gaillard J et al (2015) Inhibition of pathogenic mutant SOD1 aggregation in cultured motor neuronal cells by prevention of its SUMOylation on lysine 75. Neurodegener Dis. https://doi.org/10.1159/000439254

    Article  PubMed  Google Scholar 

  18. Oh S-M, Liu Z, Okada M, Jang SW, Liu X, Chan CB, Luo H, Ye K (2010) Ebp1 sumoylation, regulated by TLS/FUS E3 ligase, is required for its anti-proliferative activity. Oncogene 29:1017–1030. https://doi.org/10.1038/onc.2009.411

    CAS  Article  PubMed  Google Scholar 

  19. Taylor JP, Brown RH, Cleveland DW (2016) Decoding ALS: from genes to mechanism. Nature 539:197–206. https://doi.org/10.1038/nature20413

    Article  PubMed  PubMed Central  Google Scholar 

  20. Fukuda I, Ito A, Hirai G, Nishimura S, Kawasaki H, Saitoh H, Kimura K, Sodeoka M et al (2009) Ginkgolic acid inhibits protein SUMOylation by blocking formation of the E1-SUMO intermediate. Chem Biol 16:133–140. https://doi.org/10.1016/j.chembiol.2009.01.009

    CAS  Article  PubMed  Google Scholar 

  21. Sun Y, Jiang X, Chen S, Price BD (2006) Inhibition of histone acetyltransferase activity by anacardic acid sensitizes tumor cells to ionizing radiation. FEBS Lett 580:4353–4356. https://doi.org/10.1016/j.febslet.2006.06.092

    CAS  Article  PubMed  Google Scholar 

  22. Prasad A, Bharathi V, Sivalingam V, Girdhar A, Patel BK (2019) Molecular mechanisms of TDP-43 misfolding and pathology in amyotrophic lateral sclerosis. Front Mol Neurosci 12:25. https://doi.org/10.3389/fnmol.2019.00025

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. Lagier-Tourenne C, Polymenidou M, Cleveland DW (2010) TDP-43 and FUS/TLS: emerging roles in RNA processing and neurodegeneration. Hum Mol Genet 19:R46–R64. https://doi.org/10.1093/hmg/ddq137

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. Maurel C, Dangoumau A, Marouillat S, Brulard C, Chami A, Hergesheimer R, Corcia P, Blasco H et al (2018) Causative genes in amyotrophic lateral sclerosis and protein degradation pathways: a link to neurodegeneration. Mol Neurobiol 55:6480–6499. https://doi.org/10.1007/s12035-017-0856-0

    CAS  Article  PubMed  Google Scholar 

  25. Pandey D, Chen F, Patel A, Wang CY, Dimitropoulou C, Patel VS, Rudic RD, Stepp DW et al (2011) SUMO1 negatively regulates reactive oxygen species production from NADPH oxidases. Arterioscler Thromb Vasc Biol 31:1634–1642. https://doi.org/10.1161/ATVBAHA.111.226621

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  26. Gu X, Sun J, Li S, Wu X, Li L (2013) Oxidative stress induces DNA demethylation and histone acetylation in SH-SY5Y cells: potential epigenetic mechanisms in gene transcription in Aβ production. Neurobiol Aging 34:1069–1079. https://doi.org/10.1016/j.neurobiolaging.2012.10.013

    CAS  Article  PubMed  Google Scholar 

  27. Saha RN, Pahan K (2006) HATs and HDACs in neurodegeneration: a tale of disconcerted acetylation homeostasis. Cell Death Differ 13:539–550. https://doi.org/10.1038/sj.cdd.4401769

    CAS  Article  PubMed  Google Scholar 

  28. Chen B, Cepko CL (2009) HDAC4 regulates neuronal survival in normal and diseased retinas. Science 323:256–259. https://doi.org/10.1126/science.1166226

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  29. Casas C, Herrando-Grabulosa M, Manzano R, Mancuso R, Osta R, Navarro X (2013) Early presymptomatic cholinergic dysfunction in a murine model of amyotrophic lateral sclerosis. Brain Behav 3:145–158. https://doi.org/10.1002/brb3.104

    Article  PubMed  PubMed Central  Google Scholar 

  30. Bogachek MV, Park JM, De Andrade JP et al (2016) Inhibiting the SUMO pathway represses the cancer stem cell population in breast and colorectal carcinomas. Stem Cell Rep 7:1140–1151. https://doi.org/10.1016/j.stemcr.2016.11.001

    CAS  Article  Google Scholar 

  31. Casas C, Manzano R, Vaz R, Osta R, Brites D (2016) Synaptic failure: focus in an integrative view of ALS. Brain Plast Amst Neth 1:159–175. https://doi.org/10.3233/BPL-140001

    Article  Google Scholar 

  32. Andrés-Benito P, Moreno J, Aso E et al (2017) Amyotrophic lateral sclerosis, gene deregulation in the anterior horn of the spinal cord and frontal cortex area 8: implications in frontotemporal lobar degeneration. Aging 9:823–851. https://doi.org/10.18632/aging.101195

    Article  PubMed  PubMed Central  Google Scholar 

  33. Hu J-H, Zhang H, Wagey R, Krieger C, Pelech SL (2003) Protein kinase and protein phosphatase expression in amyotrophic lateral sclerosis spinal cord. J Neurochem 85:432–442. https://doi.org/10.1046/j.1471-4159.2003.01670.x

    CAS  Article  PubMed  Google Scholar 

  34. Szelechowski M, Amoedo N, Obre E, Léger C, Allard L, Bonneu M, Claverol S, Lacombe D et al (2018) Metabolic reprogramming in amyotrophic lateral sclerosis. Sci Rep 8:3953. https://doi.org/10.1038/s41598-018-22318-5

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  35. Kim HJ, Taylor JP (2017) Lost in transportation: nucleocytoplasmic transport defects in ALS and other neurodegenerative diseases. Neuron 96:285–297. https://doi.org/10.1016/j.neuron.2017.07.029

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  36. Woerner AC, Frottin F, Hornburg D, Feng LR, Meissner F, Patra M, Tatzelt J, Mann M et al (2016) Cytoplasmic protein aggregates interfere with nucleocytoplasmic transport of protein and RNA. Science 351:173–176. https://doi.org/10.1126/science.aad2033

    CAS  Article  PubMed  Google Scholar 

  37. Walker AK, Spiller KJ, Ge G, Zheng A, Xu Y, Zhou M, Tripathy K, Kwong LK et al (2015) Functional recovery in new mouse models of ALS/FTLD after clearance of pathological cytoplasmic TDP-43. Acta Neuropathol (Berl) 130:643–660. https://doi.org/10.1007/s00401-015-1460-x

    CAS  Article  Google Scholar 

  38. Zheng M, Shi Y, Fan D (2013) Nuclear TAR DNA-binding protein 43: a new target for amyotrophic lateral sclerosis treatment. Neural Regen Res 8:3284. https://doi.org/10.3969/j.issn.1673-5374.2013.35.003

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  39. Zhao X (2018) SUMO-mediated regulation of nuclear functions and signaling processes. Mol Cell 71:409–418. https://doi.org/10.1016/j.molcel.2018.07.027

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  40. Wood LD, Irvin BJ, Nucifora G, Luce KS, Hiebert SW (2003) Small ubiquitin-like modifier conjugation regulates nuclear export of TEL, a putative tumor suppressor. Proc Natl Acad Sci U S A 100:3257–3262. https://doi.org/10.1073/pnas.0637114100

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  41. Dong Y, Chen Y (2018) The role of ubiquitinated TDP-43 in amyotrophic lateral sclerosis. https://nnjournal.net/article/view/2401. Accessed 23 Jan 2019

  42. Marcelli S, Ficulle E, Piccolo L, Corbo M, Feligioni M (2018) An overview of the possible therapeutic role of SUMOylation in the treatment of Alzheimer’s disease. Pharmacol Res 130:420–437. https://doi.org/10.1016/j.phrs.2017.12.023

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgments

We thank A. Bennett for reading the manuscript. We thank the Département génomique of the plateform PST ASB, Université de Tours, for technical help. We thank Dr Frédéric Laumonnier for essential scientific advices and support towards this project.

Funding

This work was supported by a grant ARSLA; the Laboratory of Excellence (Labex) MabImprove; a PhD fellowship of the Region Centre Val-de-Loire France (CM).

Author information

Authors and Affiliations

  1. UMR 1253, iBrain, Université de Tours, Inserm, Tours, France

    Cindy Maurel, Anna A. Chami, Rose-Anne Thépault, Sylviane Marouillat, Hélène Blasco, Philippe Corcia, Christian R. Andres & Patrick Vourc’h

  2. Service de Biochimie et de Biologie Moléculaire, CHRU de Tours, 37044, Tours, France

    Hélène Blasco, Christian R. Andres & Patrick Vourc’h

  3. Service de Neurologie, CHRU de Tours, 37044, Tours, France

    Philippe Corcia

Authors
  1. Cindy Maurel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anna A. Chami
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rose-Anne Thépault
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sylviane Marouillat
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hélène Blasco
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Philippe Corcia
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christian R. Andres
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Patrick Vourc’h
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cindy Maurel.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Maurel, C., Chami, A., Thépault, RA. et al. A role for SUMOylation in the Formation and Cellular Localization of TDP-43 Aggregates in Amyotrophic Lateral Sclerosis. Mol Neurobiol 57, 1361–1373 (2020). https://doi.org/10.1007/s12035-019-01810-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12035-019-01810-7

Keywords

  • ALS
  • TDP-43
  • SUMO
  • Aggregation