Skip to main content
Springer Nature Link
Log in

PLK2 Modulates α-Synuclein Aggregation in Yeast and Mammalian Cells

  • Published:
Molecular Neurobiology Aims and scope Submit manuscript

Abstract

Phosphorylation of α-synuclein (aSyn) on serine 129 is one of the major post-translation modifications found in Lewy bodies, the typical pathological hallmark of Parkinson’s disease. Here, we found that both PLK2 and PLK3 phosphorylate aSyn on serine 129 in yeast. However, only PLK2 increased aSyn cytotoxicity and the percentage of cells presenting cytoplasmic foci. Consistently, in mammalian cells, PLK2 induced aSyn phosphorylation on serine 129 and induced an increase in the size of the inclusions. Our study supports a role for PLK2 in the generation of aSyn inclusions by a mechanism that does not depend directly on serine 129 phosphorylation.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M (1997) Alpha-synuclein in Lewy bodies. Nature 388(6645):839–840. doi:10.1038/42166

    Article  CAS  PubMed  Google Scholar 

  2. Gasser T, Hardy J, Mizuno Y (2011) Milestones in PD genetics. Mov Disord 26(6):1042–1048. doi:10.1002/mds.23637

    Article  PubMed  Google Scholar 

  3. Simon-Sanchez J, Schulte C, Bras JM, Sharma M, Gibbs JR, Berg D, Paisan-Ruiz C, Lichtner P, Scholz SW, Hernandez DG, Kruger R, Federoff M, Klein C, Goate A, Perlmutter J, Bonin M, Nalls MA, Illig T, Gieger C, Houlden H, Steffens M, Okun MS, Racette BA, Cookson MR, Foote KD, Fernandez HH, Traynor BJ, Schreiber S, Arepalli S, Zonozi R, Gwinn K, van der Brug M, Lopez G, Chanock SJ, Schatzkin A, Park Y, Hollenbeck A, Gao J, Huang X, Wood NW, Lorenz D, Deuschl G, Chen H, Riess O, Hardy JA, Singleton AB, Gasser T (2009) Genome-wide association study reveals genetic risk underlying Parkinson's disease. Nat Genet 41(12):1308–1312. doi:10.1038/ng.487

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Anderson JP, Walker DE, Goldstein JM, de Laat R, Banducci K, Caccavello RJ, Barbour R, Huang J, Kling K, Lee M, Diep L, Keim PS, Shen X, Chataway T, Schlossmacher MG, Seubert P, Schenk D, Sinha S, Gai WP, Chilcote TJ (2006) Phosphorylation of Ser-129 is the dominant pathological modification of alpha-synuclein in familial and sporadic Lewy body disease. J Biol Chem 281(40):29739–29752. doi:10.1074/jbc.M600933200

    Article  CAS  PubMed  Google Scholar 

  5. Okochi M, Walter J, Koyama A, Nakajo S, Baba M, Iwatsubo T, Meijer L, Kahle PJ, Haass C (2000) Constitutive phosphorylation of the Parkinson’s disease associated alpha-synuclein. J Biol Chem 275(1):390–397

    Article  CAS  PubMed  Google Scholar 

  6. Hasegawa M, Fujiwara H, Nonaka T, Wakabayashi K, Takahashi H, Lee VM, Trojanowski JQ, Mann D, Iwatsubo T (2002) Phosphorylated alpha-synuclein is ubiquitinated in alpha-synucleinopathy lesions. J Biol Chem 277(50):49071–49076. doi:10.1074/jbc.M208046200

    Article  CAS  PubMed  Google Scholar 

  7. Paleologou KE, Schmid AW, Rospigliosi CC, Kim HY, Lamberto GR, Fredenburg RA, Lansbury PT Jr, Fernandez CO, Eliezer D, Zweckstetter M, Lashuel HA (2008) Phosphorylation at Ser-129 but not the phosphomimics S129E/D inhibits the fibrillation of alpha-synuclein. J Biol Chem 283(24):16895–16905. doi:10.1074/jbc.M800747200

    Article  CAS  PubMed  Google Scholar 

  8. Chen L, Feany MB (2005) Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci 8(5):657–663. doi:10.1038/nn1443

    Article  CAS  PubMed  Google Scholar 

  9. Freichel C, Neumann M, Ballard T, Muller V, Woolley M, Ozmen L, Borroni E, Kretzschmar HA, Haass C, Spooren W, Kahle PJ (2007) Age-dependent cognitive decline and amygdala pathology in alpha-synuclein transgenic mice. Neurobiol Aging 28(9):1421–1435. doi:10.1016/j.neurobiolaging.2006.06.013

    Article  CAS  PubMed  Google Scholar 

  10. Gorbatyuk OS, Li S, Sullivan LF, Chen W, Kondrikova G, Manfredsson FP, Mandel RJ, Muzyczka N (2008) The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease. Proc Natl Acad Sci USA 105(2):763–768. doi:10.1073/pnas.0711053105

    Article  CAS  PubMed  Google Scholar 

  11. Azeredo da Silveira S, Schneider BL, Cifuentes-Diaz C, Sage D, Abbas-Terki T, Iwatsubo T, Unser M, Aebischer P (2009) Phosphorylation does not prompt, nor prevent, the formation of alpha-synuclein toxic species in a rat model of Parkinson's disease. Hum Mol Genet 18(5):872–887. doi:10.1093/hmg/ddn417

    CAS  PubMed  Google Scholar 

  12. McFarland NR, Fan Z, Xu K, Schwarzschild MA, Feany MB, Hyman BT, McLean PJ (2009) Alpha-synuclein S129 phosphorylation mutants do not alter nigrostriatal toxicity in a rat model of Parkinson disease. J Neuropathol Exp Neurol 68(5):515–524. doi:10.1097/NEN.0b013e3181a24b53

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Sakamoto M, Arawaka S, Hara S, Sato H, Cui C, Machiya Y, Koyama S, Wada M, Kawanami T, Kurita K, Kato T (2009) Contribution of endogenous G-protein-coupled receptor kinases to Ser129 phosphorylation of alpha-synuclein in HEK293 cells. Biochem Biophys Res Commun 384(3):378–382. doi:10.1016/j.bbrc.2009.04.130

    Article  CAS  PubMed  Google Scholar 

  14. Nakajo S, Tsukada K, Omata K, Nakamura Y, Nakaya K (1993) A new brain-specific 14-kDa protein is a phosphoprotein. Its complete amino acid sequence and evidence for phosphorylation. Eur J Biochem 217(3):1057–1063

    Article  CAS  PubMed  Google Scholar 

  15. Inglis KJ, Chereau D, Brigham EF, Chiou SS, Schobel S, Frigon NL, Yu M, Caccavello RJ, Nelson S, Motter R, Wright S, Chian D, Santiago P, Soriano F, Ramos C, Powell K, Goldstein JM, Babcock M, Yednock T, Bard F, Basi GS, Sham H, Chilcote TJ, McConlogue L, Griswold-Prenner I, Anderson JP (2009) Polo-like kinase 2 (PLK2) phosphorylates alpha-synuclein at serine 129 in central nervous system. J Biol Chem 284(5):2598–2602. doi:10.1074/jbc.C800206200

    Article  CAS  PubMed  Google Scholar 

  16. Mbefo MK, Paleologou KE, Boucharaba A, Oueslati A, Schell H, Fournier M, Olschewski D, Yin G, Zweckstetter M, Masliah E, Kahle PJ, Hirling H, Lashuel HA (2010) Phosphorylation of synucleins by members of the Polo-like kinase family. J Biol Chem 285(4):2807–2822. doi:10.1074/jbc.M109.081950

    Article  CAS  PubMed  Google Scholar 

  17. Waxman EA, Giasson BI (2011) Characterization of kinases involved in the phosphorylation of aggregated alpha-synuclein. J Neurosci Res 89(2):231–247. doi:10.1002/jnr.22537

    Article  CAS  PubMed  Google Scholar 

  18. Gitler AD, Bevis BJ, Shorter J, Strathearn KE, Hamamichi S, Su LJ, Caldwell KA, Caldwell GA, Rochet JC, McCaffery JM, Barlowe C, Lindquist S (2008) The Parkinson's disease protein alpha-synuclein disrupts cellular Rab homeostasis. Proc Natl Acad Sci USA 105(1):145–150. doi:10.1073/pnas.0710685105

    Article  CAS  PubMed  Google Scholar 

  19. Outeiro TF, Lindquist S (2003) Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science 302(5651):1772–1775. doi:10.1126/science.1090439

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  20. Tenreiro S, Outeiro TF (2010) Simple is good: yeast models of neurodegeneration. FEMS Yeast Res 10(8):970–979. doi:10.1111/j.1567-1364.2010.00649.x

    Article  CAS  PubMed  Google Scholar 

  21. Visanji NP, Wislet-Gendebien S, Oschipok LW, Zhang G, Aubert I, Fraser PE, Tandon A (2011) Effect of Ser-129 phosphorylation on interaction of alpha-synuclein with synaptic and cellular membranes. J Biol Chem 286(41):35863–35873. doi:10.1074/jbc.M111.253450

    Article  CAS  PubMed  Google Scholar 

  22. Outeiro TF, Putcha P, Tetzlaff JE, Spoelgen R, Koker M, Carvalho F, Hyman BT, McLean PJ (2008) Formation of toxic oligomeric alpha-synuclein species in living cells. PLoS One 3(4):e1867. doi:10.1371/journal.pone.0001867

    Article  PubMed Central  PubMed  Google Scholar 

  23. McLean PJ, Kawamata H, Hyman BT (2001) Alpha-synuclein-enhanced green fluorescent protein fusion proteins form proteasome sensitive inclusions in primary neurons. Neuroscience 104(3):901–912

    Article  CAS  PubMed  Google Scholar 

  24. Engelender S, Kaminsky Z, Guo X, Sharp AH, Amaravi RK, Kleiderlein JJ, Margolis RL, Troncoso JC, Lanahan AA, Worley PF, Dawson VL, Dawson TM, Ross CA (1999) Synphilin-1 associates with alpha-synuclein and promotes the formation of cytosolic inclusions. Nat Genet 22(1):110–114. doi:10.1038/8820

    CAS  PubMed  Google Scholar 

  25. Wakabayashi K, Engelender S, Yoshimoto M, Tsuji S, Ross CA, Takahashi H (2000) Synphilin-1 is present in Lewy bodies in Parkinson’s disease. Ann Neurol 47(4):521–523

    CAS  PubMed  Google Scholar 

  26. Chung KK, Zhang Y, Lim KL, Tanaka Y, Huang H, Gao J, Ross CA, Dawson VL, Dawson TM (2001) Parkin ubiquitinates the alpha-synuclein-interacting protein, synphilin-1: implications for Lewy-body formation in Parkinson disease. Nat Med 7(10):1144–1150. doi:10.1038/nm1001-1144 nm1001-1144

    Article  CAS  PubMed  Google Scholar 

  27. Lim KL, Chew KC, Tan JM, Wang C, Chung KK, Zhang Y, Tanaka Y, Smith W, Engelender S, Ross CA, Dawson VL, Dawson TM (2005) Parkin mediates nonclassical, proteasomal-independent ubiquitination of synphilin-1: implications for Lewy body formation. J Neurosci 25(8):2002–2009. doi:10.1523/JNEUROSCI.4474-04.2005

    Article  CAS  PubMed  Google Scholar 

  28. Lee G, Tanaka M, Park K, Lee SS, Kim YM, Junn E, Lee SH, Mouradian MM (2004) Casein kinase II-mediated phosphorylation regulates alpha-synuclein/synphilin-1 interaction and inclusion body formation. J Biol Chem 279(8):6834–6839. doi:10.1074/jbc.M312760200

    Article  CAS  PubMed  Google Scholar 

  29. Avraham E, Szargel R, Eyal A, Rott R, Engelender S (2005) Glycogen synthase kinase 3beta modulates synphilin-1 ubiquitylation and cellular inclusion formation by SIAH: implications for proteasomal function and Lewy body formation. J Biol Chem 280(52):42877–42886. doi:10.1074/jbc.M505608200

    Article  CAS  PubMed  Google Scholar 

  30. Smith WW, Margolis RL, Li X, Troncoso JC, Lee MK, Dawson VL, Dawson TM, Iwatsubo T, Ross CA (2005) Alpha-synuclein phosphorylation enhances eosinophilic cytoplasmic inclusion formation in SH-SY5Y cells. J Neurosci 25(23):5544–5552. doi:10.1523/JNEUROSCI.0482-05.2005

    Article  CAS  PubMed  Google Scholar 

  31. Wang S, Xu B, Liou LC, Ren Q, Huang S, Luo Y, Zhang Z, Witt SN Alpha-synuclein disrupts stress signaling by inhibiting polo-like kinase Cdc5/Plk2. Proc Natl Acad Sci USA 109 (40):16119-16124. doi:1206286109 [pii]10.1073/pnas.1206286109

  32. Matthew EM, Hart LS, Astrinidis A, Navaraj A, Dolloff NG, Dicker DT, Henske EP, El-Deiry WS (2009) The p53 target Plk2 interacts with TSC proteins impacting mTOR signaling, tumor growth and chemosensitivity under hypoxic conditions. Cell Cycle 8(24):4168–4175

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  33. Webb JL, Ravikumar B, Atkins J, Skepper JN, Rubinsztein DC (2003) Alpha-synuclein is degraded by both autophagy and the proteasome. J Biol Chem 278(27):25009–25013. doi:10.1074/jbc.M300227200

    Article  CAS  PubMed  Google Scholar 

  34. Tanese N (1997) Small-scale density gradient sedimentation to separate and analyze multiprotein complexes. Methods 12(3):224–234. doi:10.1006/meth.1997.0475

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Fundação para a Ciência e Tecnologia (PTDC/SAU-NEU/105215/2008 and PTDC/BIA-BCM/117975/2010) [SFRH/BPD/65890/2009 to Z.M., SFRH/BPD/35767/2007 to S.T., SFRH/BI/5177/2011 to P.A, SFRH/BD/79337/2011 to S.G]. TFO was supported by a Marie Curie International Reintegration Grant and an EMBO Installation Grant. EB is supported by EC Framework 7 Marie Curie Fellowship Training Network Grant (NEURASYNC) and by EU FP7 MEFOPA. SG was supported by AXA Research Fund.

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Cell and Molecular Neuroscience Unit, Instituto de Medicina Molecular, Lisbon, Portugal

    Elisa Basso, Pedro Antas, Zrinka Marijanovic, Susana Gonçalves, Sandra Tenreiro & Tiago Fleming Outeiro

  2. Instituto de Fisiologia, Faculdade de Medicina da Universidade de Lisboa, Lisbon, Portugal

    Tiago Fleming Outeiro

  3. Department of Neurodegeneration and Restorative Research, Center for Nanoscale Microscopy and Molecular Physiology of the Brain, University Medical Center Göttingen, Göttingen, Germany

    Tiago Fleming Outeiro

Authors
  1. Elisa Basso
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pedro Antas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zrinka Marijanovic
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Susana Gonçalves
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sandra Tenreiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tiago Fleming Outeiro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tiago Fleming Outeiro.

Additional information

Elisa Basso and Pedro Antas have equal contribution.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Fig. S1

PLK4 does not increase aSyn toxicity and foci formation. a Spotting assay of yeast cultures co-expressing aSyn (aSyn + EV) or empty vector (EV) with PLK4. The yeast cell suspensions were serial diluted (1/2) and applied as spots (4 μl) onto the surface of the solid medium either with glucose (control) or galactose (induced of aSyn and PLK4 expression) as carbon source. b Fluorescence microscopy and quantification of the number of cells presenting aSyn foci in cells expressing aSyn-GFP fusion protein alone (aSyn-GFP + EV) or together with PLK4. (JPEG 2223 kb)

High resolution image (EPS 18683 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Basso, E., Antas, P., Marijanovic, Z. et al. PLK2 Modulates α-Synuclein Aggregation in Yeast and Mammalian Cells. Mol Neurobiol 48, 854–862 (2013). https://doi.org/10.1007/s12035-013-8473-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12035-013-8473-z

Keywords

Search

Navigation