Skip to main content
SpringerLink
Log in

Inhibition of Autophagy via p53-Mediated Disruption of ULK1 in a SCA7 Polyglutamine Disease Model

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Spinocerebellar ataxia type 7 (SCA7) is one of nine neurodegenerative disorders caused by expanded polyglutamine domains. These so-called polyglutamine (polyQ) diseases are all characterized by aggregation. Reducing the level of aggregating polyQ proteins via pharmacological activation of autophagy has been suggested as a therapeutic approach. However, recently, evidence implicating autophagic dysfunction in these disorders has also been reported. In this study, we show that the SCA7 polyglutamine protein ataxin-7 (ATXN7) reduces the autophagic activity via a previously unreported mechanism involving p53-mediated disruption of two key proteins involved in autophagy initiation. We show that in mutant ATXN7 cells, an increased p53–FIP200 interaction and co-aggregation of p53–FIP200 into ATXN7 aggregates result in decreased soluble FIP200 levels and subsequent destabilization of ULK1. Together, this leads to a decreased capacity for autophagy induction via the ULK1–FIP200–Atg13–Atg101 complex. We also show that treatment with a p53 inhibitor, or a blocker of ATXN7 aggregation, can restore the soluble levels of FIP200 and ULK1, as well as increase the autophagic activity and reduce ATXN7 toxicity. Understanding the mechanism behind polyQ-mediated inhibition of autophagy is of importance if therapeutic approaches based on autophagy stimulation should be developed for these disorders.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

Abbreviations

Atg:

Autophagy-related gene

ATXN7:

Ataxin-7

BafA:

Bafilomycin A1

GFP:

Green fluorescence protein

HD:

Hungtinton’s disease

kDa:

Kilodalton

PolyQ:

Polyglutamine

Q:

Glutamine

QBP1:

Polyglutamine binding peptide 1

SCA7:

Spinocerebellar ataxia type 7

References

  • Ajayi A, Yu X, Lindberg S, Langel U, Strom AL (2012) Expanded ataxin-7 cause toxicity by inducing ROS production from NADPH oxidase complexes in a stable inducible spinocerebellar ataxia type 7 (SCA7) model. BMC Neurosci 13:86

    Article  PubMed  CAS  Google Scholar 

  • Bae BI, Xu H, Igarashi S, Fujimuro M, Agrawal N, Taya Y, Hayward SD, Moran TH, Montell C, Ross CA, Snyder SH, Sawa A (2005) p53 mediates cellular dysfunction and behavioral abnormalities in Huntington's disease. Neuron 47:29–41

    Article  PubMed  CAS  Google Scholar 

  • Chen Y, Klionsky DJ (2011) The regulation of autophagy—unanswered questions. J Cell Sci 124:161–170

    Article  PubMed  CAS  Google Scholar 

  • Cheung ZH, Ip NY (2011) Autophagy deregulation in neurodegenerative diseases—recent advances and future perspectives. J Neurochem 118:317–325

    Article  PubMed  CAS  Google Scholar 

  • Chou AH, Lin AC, Hong KY, Hu SH, Chen YL, Chen JY, Wang HL (2011) p53 activation mediates polyglutamine-expanded ataxin-3 upregulation of Bax expression in cerebellar and pontine nuclei neurons. Neurochem Int 58:145–152

    Article  PubMed  CAS  Google Scholar 

  • Garden GA, Libby RT, Fu YH, Kinoshita Y, Huang J, Possin DE, Smith AC, Martinez RA, Fine GC, Grote SK, Ware CB, Einum DD, Morrison RS, Ptacek LJ, Sopher BL, La Spada AR (2002) Polyglutamine-expanded ataxin-7 promotes non-cell-autonomous Purkinje cell degeneration and displays proteolytic cleavage in ataxic transgenic mice. J Neurosci 22:4897–4905

    PubMed  CAS  Google Scholar 

  • Grison A, Mantovani F, Comel A, Agostoni E, Gustincich S, Persichetti F, Del Sal G (2011) Ser46 phosphorylation and prolyl-isomerase Pin1-mediated isomerization of p53 are key events in p53-dependent apoptosis induced by mutant huntingtin. Proc Natl Acad Sci U S A 108:17979–17984

    Article  PubMed  CAS  Google Scholar 

  • Hara T, Nakamura K, Matsui M, Yamamoto A, Nakahara Y, Suzuki-Migishima R, Yokoyama M, Mishima K, Saito I, Okano H, Mizushima N (2006) Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature 441:885–889

    Article  PubMed  CAS  Google Scholar 

  • Hara T, Takamura A, Kishi C, Iemura S, Natsume T, Guan JL, Mizushima N (2008) FIP200, a ULK-interacting protein, is required for autophagosome formation in mammalian cells. J Cell Biol 181:497–510

    Article  PubMed  CAS  Google Scholar 

  • He C, Klionsky DJ (2009) Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet 43:67–93

    Article  PubMed  CAS  Google Scholar 

  • Helmlinger D, Hardy S, Sasorith S, Klein F, Robert F, Weber C, Miguet L, Potier N, Van-Dorsselaer A, Wurtz JM, Mandel JL, Tora L, Devys D (2004) Ataxin-7 is a subunit of GCN5 histone acetyltransferase-containing complexes. Hum Mol Genet 13:1257–1265

    Article  PubMed  CAS  Google Scholar 

  • Heng MY, Duong DK, Albin RL, Tallaksen-Greene SJ, Hunter JM, Lesort MJ, Osmand A, Paulson HL, Detloff PJ (2010) Early autophagic response in a novel knock-in model of Huntington disease. Hum Mol Genet 19:3702–3720

    Article  PubMed  CAS  Google Scholar 

  • Illuzzi JL, Vickers CA, Kmiec EB (2011) Modifications of p53 and the DNA damage response in cells expressing mutant form of the protein huntingtin. J Mol Neurosci 45:256–268

    Article  PubMed  CAS  Google Scholar 

  • Jonasson J, Strom AL, Hart P, Brannstrom T, Forsgren L, Holmberg M (2002) Expression of ataxin-7 in CNS and non-CNS tissue of normal and SCA7 individuals. Acta Neuropathol 104:29–37

    Article  PubMed  CAS  Google Scholar 

  • Katsuno M, Banno H, Suzuki K, Takeuchi Y, Kawashima M, Tanaka F, Adachi H, Sobue G (2008) Molecular genetics and biomarkers of polyglutamine diseases. Curr Mol Med 8:221–234

    Article  PubMed  CAS  Google Scholar 

  • Klionsky DJ, Abdalla FC, Abeliovich H, Abraham RT, Acevedo-Arozena A, Adeli K, Agholme L, Agnello M, Agostinis P, Aguirre-Ghiso JA, Ahn HJ, Ait-Mohamed O, Ait-Si-Ali S, Akematsu T, Akira S, Al-Younes HM, Al-Zeer MA, Albert ML, Albin RL, Alegre-Abarrategui J, Aleo MF, Alirezaei M, Almasan A, Almonte-Becerril M, Amano A, Amaravadi R, Amarnath S, Amer AO, Andrieu-Abadie N, Anantharam V, Ann DK, Anoopkumar-Dukie S, Aoki H, Apostolova N, Arancia G, Aris JP, Asanuma K, Asare NY, Ashida H, Askanas V, Askew DS, Auberger P, Baba M, Backues SK, Baehrecke EH, Bahr BA, Bai XY, Bailly Y, Baiocchi R, Baldini G, Balduini W, Ballabio A, Bamber BA, Bampton ET, Banhegyi G, Bartholomew CR, Bassham DC, Bast RC Jr, Batoko H, Bay BH, Beau I, Bechet DM, Begley TJ, Behl C, Behrends C, Bekri S, Bellaire B, Bendall LJ, Benetti L, Berliocchi L, Bernardi H, Bernassola F, Besteiro S, Bhatia-Kissova I, Bi X, Biard-Piechaczyk M, Blum JS, Boise LH, Bonaldo P, Boone DL, Bornhauser BC, Bortoluci KR, Bossis I, Bost F, Bourquin JP, Boya P, Boyer-Guittaut M, Bozhkov PV, Brady NR, Brancolini C, Brech A, Brenman JE, Brennand A, Bresnick EH, Brest P, Bridges D, Bristol ML, Brookes PS, Brown EJ, Brumell JH et al (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8:445–544

    Article  PubMed  CAS  Google Scholar 

  • Komatsu M, Wang QJ, Holstein GR, Friedrich VL Jr, Iwata J, Kominami E, Chait BT, Tanaka K, Yue Z (2007) Essential role for autophagy protein Atg7 in the maintenance of axonal homeostasis and the prevention of axonal degeneration. Proc Natl Acad Sci U S A 104:14489–14494

    Article  PubMed  CAS  Google Scholar 

  • Konigsmark BW, Weiner LP (1970) The olivopontocerebellar atrophies: a review. Medicine (Baltimore) 49:227–241

    CAS  Google Scholar 

  • Li X, Li H, Li XJ (2008) Intracellular degradation of misfolded proteins in polyglutamine neurodegenerative diseases. Brain Res Rev 59:245–252

    Article  PubMed  CAS  Google Scholar 

  • Liang CC, Wang C, Peng X, Gan B, Guan JL (2010) Neural-specific deletion of FIP200 leads to cerebellar degeneration caused by increased neuronal death and axon degeneration. J Biol Chem 285:3499–3509

    Article  PubMed  CAS  Google Scholar 

  • Lindenberg KS, Yvert G, Muller K, Landwehrmeyer GB (2000) Expression analysis of ataxin-7 mRNA and protein in human brain: evidence for a widespread distribution and focal protein accumulation. Brain Pathol 10:385–394

    Article  PubMed  CAS  Google Scholar 

  • Maiuri MC, Galluzzi L, Morselli E, Kepp O, Malik SA, Kroemer G (2010) Autophagy regulation by p53. Curr Opin Cell Biol 22:181–185

    Article  PubMed  CAS  Google Scholar 

  • Martin JJ, Van Regemorter N, Krols L, Brucher JM, de Barsy T, Szliwowski H, Evrard P, Ceuterick C, Tassignon MJ, Smet-Dieleman H et al (1994) On an autosomal dominant form of retinal-cerebellar degeneration: an autopsy study of five patients in one family. Acta Neuropathol 88:277–286

    Article  PubMed  CAS  Google Scholar 

  • Martinez-Vicente M, Talloczy Z, Wong E, Tang G, Koga H, Kaushik S, de Vries R, Arias E, Harris S, Sulzer D, Cuervo AM (2010) Cargo recognition failure is responsible for inefficient autophagy in Huntington's disease. Nat Neurosci 13:567–576

    Article  PubMed  CAS  Google Scholar 

  • Metzger S, Saukko M, Van Che H, Tong L, Puder Y, Riess O, Nguyen HP (2010) Age at onset in Huntington's disease is modified by the autophagy pathway: implication of the V471A polymorphism in Atg7. Hum Genet 128:453–459

    Article  PubMed  CAS  Google Scholar 

  • Mizushima N (2010) The role of the Atg1/ULK1 complex in autophagy regulation. Curr Opin Cell Biol 22:132–139

    Article  PubMed  CAS  Google Scholar 

  • Morselli E, Shen S, Ruckenstuhl C, Bauer MA, Marino G, Galluzzi L, Criollo A, Michaud M, Maiuri MC, Chano T, Madeo F, Kroemer G (2011) p53 inhibits autophagy by interacting with the human ortholog of yeast Atg17, RB1CC1/FIP200. Cell Cycle 10:2763–2769

    Article  PubMed  CAS  Google Scholar 

  • Morselli E, Tasdemir E, Maiuri MC, Galluzzi L, Kepp O, Criollo A, Vicencio JM, Soussi T, Kroemer G (2008) Mutant p53 protein localized in the cytoplasm inhibits autophagy. Cell Cycle 7:3056–3061

    Article  PubMed  CAS  Google Scholar 

  • Nagai Y, Fujikake N, Ohno K, Higashiyama H, Popiel HA, Rahadian J, Yamaguchi M, Strittmatter WJ, Burke JR, Toda T (2003) Prevention of polyglutamine oligomerization and neurodegeneration by the peptide inhibitor QBP1 in Drosophila. Hum Mol Genet 12:1253–1259

    Article  PubMed  CAS  Google Scholar 

  • Nagai Y, Tucker T, Ren H, Kenan DJ, Henderson BS, Keene JD, Strittmatter WJ, Burke JR (2000) Inhibition of polyglutamine protein aggregation and cell death by novel peptides identified by phage display screening. J Biol Chem 275:10437–10442

    Article  PubMed  CAS  Google Scholar 

  • Nakamura S, Roth JA, Mukhopadhyay T (2000) Multiple lysine mutations in the C-terminal domain of p53 interfere with MDM2-dependent protein degradation and ubiquitination. Mol Cell Biol 20:9391–9398

    Article  PubMed  CAS  Google Scholar 

  • Nakamura Y, Tagawa K, Oka T, Sasabe T, Ito H, Shiwaku H, La Spada AR, Okazawa H (2012) Ataxin-7 associates with microtubules and stabilizes the cytoskeletal network. Hum Mol Genet 21:1099–1110

    Article  PubMed  CAS  Google Scholar 

  • Palhan VB, Chen S, Peng GH, Tjernberg A, Gamper AM, Fan Y, Chait BT, La Spada AR, Roeder RG (2005) Polyglutamine-expanded ataxin-7 inhibits STAGA histone acetyltransferase activity to produce retinal degeneration. Proc Natl Acad Sci U S A 102:8472–8477

    Article  PubMed  CAS  Google Scholar 

  • Qin Q, Inatome R, Hotta A, Kojima M, Yamamura H, Hirai H, Yoshizawa T, Tanaka H, Fukami K, Yanagi S (2006) A novel GTPase, CRAG, mediates promyelocytic leukemia protein-associated nuclear body formation and degradation of expanded polyglutamine protein. J Cell Biol 172:497–504

    Article  PubMed  CAS  Google Scholar 

  • Ravikumar B, Duden R, Rubinsztein DC (2002) Aggregate-prone proteins with polyglutamine and polyalanine expansions are degraded by autophagy. Hum Mol Genet 11:1107–1117

    Article  PubMed  CAS  Google Scholar 

  • Ravikumar B, Vacher C, Berger Z, Davies JE, Luo S, Oroz LG, Scaravilli F, Easton DF, Duden R, O'Kane CJ, Rubinsztein DC (2004) Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease. Nat Genet 36:585–595

    Article  PubMed  CAS  Google Scholar 

  • Shahbazian MD, Orr HT, Zoghbi HY (2001) Reduction of Purkinje cell pathology in SCA1 transgenic mice by p53 deletion. Neurobiol Dis 8:974–981

    Article  PubMed  CAS  Google Scholar 

  • Steffan JS, Kazantsev A, Spasic-Boskovic O, Greenwald M, Zhu YZ, Gohler H, Wanker EE, Bates GP, Housman DE, Thompson LM (2000) The Huntington's disease protein interacts with p53 and CREB-binding protein and represses transcription. Proc Natl Acad Sci U S A 97:6763–6768

    Article  PubMed  CAS  Google Scholar 

  • Strom AL, Forsgren L, Holmberg M (2005) A role for both wild-type and expanded ataxin-7 in transcriptional regulation. Neurobiol Dis 20:646–655

    Article  PubMed  Google Scholar 

  • Tanaka M, Machida Y, Niu S, Ikeda T, Jana NR, Doi H, Kurosawa M, Nekooki M, Nukina N (2004) Trehalose alleviates polyglutamine-mediated pathology in a mouse model of Huntington disease. Nat Med 10:148–154

    Article  PubMed  CAS  Google Scholar 

  • Tasdemir E, Maiuri MC, Galluzzi L, Vitale I, Djavaheri-Mergny M, D'Amelio M, Criollo A, Morselli E, Zhu C, Harper F, Nannmark U, Samara C, Pinton P, Vicencio JM, Carnuccio R, Moll UM, Madeo F, Paterlini-Brechot P, Rizzuto R, Szabadkai G, Pierron G, Blomgren K, Tavernarakis N, Codogno P, Cecconi F, Kroemer G (2008) Regulation of autophagy by cytoplasmic p53. Nat Cell Biol 10:676–687

    Article  PubMed  CAS  Google Scholar 

  • Tsoi H, Lau TC, Tsang SY, Lau KF, Chan HY (2012) CAG expansion induces nucleolar stress in polyglutamine diseases. Proc Natl Acad Sci U S A 109:13428–13433

    Article  PubMed  CAS  Google Scholar 

  • Wang HL, Chou AH, Lin AC, Chen SY, Weng YH, Yeh TH (2010) Polyglutamine-expanded ataxin-7 upregulates Bax expression by activating p53 in cerebellar and inferior olivary neurons. Exp Neurol 224:486–494

    Article  PubMed  CAS  Google Scholar 

  • Wang T, Lao U, Edgar BA (2009) TOR-mediated autophagy regulates cell death in Drosophila neurodegenerative disease. J Cell Biol 186:703–711

    Article  PubMed  CAS  Google Scholar 

  • Wong E, Cuervo AM (2010) Autophagy gone awry in neurodegenerative diseases. Nat Neurosci 13:805–811

    Article  PubMed  CAS  Google Scholar 

  • Young JE, Gouw L, Propp S, Sopher BL, Taylor J, Lin A, Hermel E, Logvinova A, Chen SF, Chen S, Bredesen DE, Truant R, Ptacek LJ, La Spada AR, Ellerby LM (2007) Proteolytic cleavage of ataxin-7 by caspase-7 modulates cellular toxicity and transcriptional dysregulation. J Biol Chem 282:30150–30160

    Article  PubMed  CAS  Google Scholar 

  • Young JE, Martinez RA, La Spada AR (2009) Nutrient deprivation induces neuronal autophagy and implicates reduced insulin signaling in neuroprotective autophagy activation. J Biol Chem 284:2363–2373

    Article  PubMed  CAS  Google Scholar 

  • Yu X, Ajayi A, Boga NR, Strom AL (2012) Differential degradation of full-length and cleaved ataxin-7 fragments in a novel stable inducible SCA7 model. J Mol Neurosci 47:219–233

    Article  PubMed  CAS  Google Scholar 

  • Yvert G, Lindenberg KS, Devys D, Helmlinger D, Landwehrmeyer GB, Mandel JL (2001) SCA7 mouse models show selective stabilization of mutant ataxin-7 and similar cellular responses in different neuronal cell types. Hum Mol Genet 10:1679–1692

    Article  PubMed  CAS  Google Scholar 

  • Zander C, Takahashi J, El Hachimi KH, Fujigasaki H, Albanese V, Lebre AS, Stevanin G, Duyckaerts C, Brice A (2001) Similarities between spinocerebellar ataxia type 7 (SCA7) cell models and human brain: proteins recruited in inclusions and activation of caspase-3. Hum Mol Genet 10:2569–2579

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Jessica Lundqvist and Christina Svensson for their help with RNA and RT-PCR analyses and Dr. Monica Holmberg, Umeå University, for ATXN7 constructs and antibodies. This work was supported by the Swedish Research Council (90274201), Harald Jeanssons stiftelse, Harald och Greta Jeanssons stiftelse, Magn Bergvalls stiftelse, O.E. och Edla Johanssons vetenskapliga stiftelse, Åhlen stiftelsen, Goljes stiftelse, and The Swedish Association of Persons with Neurological Disabilities.

Author information

Authors and Affiliations

  1. Department of Neurochemistry, Stockholm University, 106 91, Stockholm, Sweden

    Xin Yu, Andrés Muñoz-Alarcón, Abiodun Ajayi, Kristin E. Webling, Anne Steinhof, Ülo Langel & Anna-Lena Ström

Authors
  1. Xin Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrés Muñoz-Alarcón
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Abiodun Ajayi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kristin E. Webling
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Anne Steinhof
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ülo Langel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Anna-Lena Ström
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anna-Lena Ström.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 1081 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yu, X., Muñoz-Alarcón, A., Ajayi, A. et al. Inhibition of Autophagy via p53-Mediated Disruption of ULK1 in a SCA7 Polyglutamine Disease Model. J Mol Neurosci 50, 586–599 (2013). https://doi.org/10.1007/s12031-013-0012-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-013-0012-x

Keywords

Search

Navigation