Phosphorylation Modulates Clearance of Alpha-Synuclein Inclusions in a Yeast Model of Parkinson's Disease

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Protein aggregation is a common hallmark in neurodegenerative disorders, but is also associated with phenotypic plasticity in a variety of organisms, including yeasts. Alpha-synuclein (aSyn) forms aggregates that are typical of synucleinopathies, and is phosphorylated at S129, but the significance of phosphorylation in the biology and pathophysiology of the protein is still controversial. Exploring the power of budding yeast, we found phosphorylation reduced aSyn toxicity and inclusion formation. While inclusions formed by WT aSyn were homogeneous, those formed by S129A aSyn were larger and heterogeneous. Interestingly, clearance of aSyn inclusions was reduced in cells expressing S129A aSyn, correlating with deficient autophagy activation. The finding that phosphorylation alters the ability of cells to clear aSyn inclusions provides novel insight into the role phosphorylation may have in synucleinopathies, and suggests posttranslational modifications might constitute switches cells use to control the aggregation and clearance of key proteins, opening novel avenues for the development of therapeutic strategies for these devastating disorders.

Vyšlo v časopise: Phosphorylation Modulates Clearance of Alpha-Synuclein Inclusions in a Yeast Model of Parkinson's Disease. PLoS Genet 10(5): e32767. doi:10.1371/journal.pgen.1004302
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004302

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1. MalinovskaL, KroschwaldS, AlbertiS (2013) Protein disorder, prion propensities, and self-organizing macromolecular collectives. Biochim Biophys Acta 1834: 918–931.

2. DouglasPM, DillinA (2010) Protein homeostasis and aging in neurodegeneration. J Cell Biol 190: 719–729.

3. SpillantiniMG, CrowtherRA, JakesR, CairnsNJ, LantosPL, et al. (1998) Filamentous alpha-synuclein inclusions link multiple system atrophy with Parkinson's disease and dementia with Lewy bodies. Neurosci Lett 251: 205–208.

4. SpillantiniMG, SchmidtML, LeeVM, TrojanowskiJQ, JakesR, et al. (1997) Alpha-synuclein in Lewy bodies. Nature 388: 839–840.

5. KleinC, Lohmann-HedrichK (2007) Impact of recent genetic findings in Parkinson's disease. Curr Opin Neurol 20: 453–464.

6. AndersonJP, WalkerDE, GoldsteinJM, de LaatR, BanducciK, et al. (2006) Phosphorylation of Ser-129 is the dominant pathological modification of alpha-synuclein in familial and sporadic Lewy body disease. J Biol Chem 281: 29739–29752.

7. ChenL, FeanyMB (2005) Alpha-synuclein phosphorylation controls neurotoxicity and inclusion formation in a Drosophila model of Parkinson disease. Nat Neurosci 8: 657–663.

8. FreichelC, NeumannM, BallardT, MullerV, WoolleyM, et al. (2007) Age-dependent cognitive decline and amygdala pathology in alpha-synuclein transgenic mice. Neurobiol Aging 28: 1421–1435.

9. KuwaharaT, TonegawaR, ItoG, MitaniS, IwatsuboT (2012) Phosphorylation of alpha-synuclein protein at Ser-129 reduces neuronal dysfunction by lowering its membrane binding property in Caenorhabditis elegans. J Biol Chem 287: 7098–7109.

10. Azeredo da SilveiraS, SchneiderBL, Cifuentes-DiazC, SageD, Abbas-TerkiT, et al. (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: 872–887.

11. McFarlandNR, FanZ, XuK, SchwarzschildMA, FeanyMB, et al. (2009) Alpha-synuclein S129 phosphorylation mutants do not alter nigrostriatal toxicity in a rat model of Parkinson disease. J Neuropathol Exp Neurol 68: 515–524.

12. SmithWW, MargolisRL, LiX, TroncosoJC, LeeMK, et al. (2005) Alpha-synuclein phosphorylation enhances eosinophilic cytoplasmic inclusion formation in SH-SY5Y cells. J Neurosci 25: 5544–5552.

13. WaxmanEA, GiassonBI (2008) Specificity and regulation of casein kinase-mediated phosphorylation of alpha-synuclein. J Neuropathol Exp Neurol 67: 402–416.

14. FiskeM, ValtierraS, SolvangK, ZorniakM, WhiteM, et al. (2011) Contribution of alanine-76 and serine phosphorylation in alpha-synuclein membrane association and aggregation in yeasts. Parkinson's Disease 2011: 392180.

15. TenreiroS, MunderMC, AlbertiS, OuteiroTF (2013) Harnessing the power of yeast to unravel the molecular basis of neurodegeneration. J Neurochem 127: 438–452.

16. OuteiroTF, LindquistS (2003) Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science 302: 1772–1775.

17. GitlerAD, ChesiA, GeddieML, StrathearnKE, HamamichiS, et al. (2009) Alpha-synuclein is part of a diverse and highly conserved interaction network that includes PARK9 and manganese toxicity. Nat Genet 41: 308–315.

18. Yeger-LotemE, RivaL, SuLJ, GitlerAD, CashikarAG, et al. (2009) Bridging high-throughput genetic and transcriptional data reveals cellular responses to alpha-synuclein toxicity. Nat Genet 41: 316–323.

19. ButtnerS, BittoA, RingJ, AugstenM, ZabrockiP, et al. (2008) Functional mitochondria are required for alpha-synuclein toxicity in aging yeast. J Biol Chem 283: 7554–7560.

20. SuLJ, AuluckPK, OuteiroTF, Yeger-LotemE, KritzerJA, et al. (2010) Compounds from an unbiased chemical screen reverse both ER-to-Golgi trafficking defects and mitochondrial dysfunction in Parkinson's disease models. Dis Model Mech 3: 194–208.

21. SharmaN, BrandisKA, HerreraSK, JohnsonBE, VaidyaT, et al. (2006) Alpha-synuclein budding yeast model: toxicity enhanced by impaired proteasome and oxidative stress. J Mol Neurosci 28: 161–178.

22. ChenQ, ThorpeJ, KellerJN (2005) Alpha-synuclein alters proteasome function, protein synthesis, and stationary phase viability. J Biol Chem 280: 30009–30017.

23. WittSN, FlowerTR (2006) Alpha-synuclein, oxidative stress and apoptosis from the perspective of a yeast model of Parkinson's disease. FEMS Yeast Res 6: 1107–1116.

24. Sampaio-MarquesB, FelgueirasC, SilvaA, RodriguesM, TenreiroS, et al. (2012) SNCA (alpha-synuclein)-induced toxicity in yeast cells is dependent on sirtuin 2 (Sir2)-mediated mitophagy. Autophagy 8: 1494–1509.

25. PetroiD, PopovaB, Taheri-TaleshN, IrnigerS, ShahpasandzadehH, et al. (2012) Aggregate clearance of alpha-synuclein in Saccharomyces cerevisiae depends more on autophagosome and vacuole function than on the proteasome. J Biol Chem 287: 27567–27579.

26. SoperJH, RoyS, StieberA, LeeE, WilsonRB, et al. (2008) Alpha-synuclein-induced aggregation of cytoplasmic vesicles in Saccharomyces cerevisiae. Mol Biol Cell 19: 1093–1103.

27. GitlerAD, BevisBJ, ShorterJ, StrathearnKE, HamamichiS, et al. (2008) The Parkinson's disease protein alpha-synuclein disrupts cellular Rab homeostasis. Proc Natl Acad Sci U S A 105: 145–150.

28. CooperAA, GitlerAD, CashikarA, HaynesCM, HillKJ, et al. (2006) Alpha-synuclein blocks ER-Golgi traffic and Rab1 rescues neuron loss in Parkinson's models. Science 313: 324–328.

29. CookC, StetlerC, PetrucelliL (2012) Disruption of protein quality control in Parkinson's disease. Cold Spring Harb Perspect Biol 2: a009423.

30. XilouriM, VogiatziT, VekrellisK, ParkD, StefanisL (2009) Abberant alpha-synuclein confers toxicity to neurons in part through inhibition of chaperone-mediated autophagy. PLoS ONE 4: e5515.

31. IwataA, MaruyamaM, AkagiT, HashikawaT, KanazawaI, et al. (2003) Alpha-synuclein degradation by serine protease neurosin: implication for pathogenesis of synucleinopathies. Hum Mol Genet 12: 2625–2635.

32. LimKL, TanJM (2007) Role of the ubiquitin proteasome system in Parkinson's disease. BMC Biochemistry 8 Suppl 1S13.

33. CuervoAM, StefanisL, FredenburgR, LansburyPT, SulzerD (2004) Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy. Science 305: 1292–1295.

34. ZhangNY, TangZ, LiuCW (2008) Alpha-synuclein protofibrils inhibit 26 S proteasome-mediated protein degradation: understanding the cytotoxicity of protein protofibrils in neurodegenerative disease pathogenesis. J Biol Chem 283: 20288–20298.

35. ChuCT (2011) Diversity in the regulation of autophagy and mitophagy: lessons from Parkinson's disease. Parkinson's Disease 2011: 789431.

36. KraghCL, UbhiK, Wyss-CorayT, MasliahE (2012) Autophagy in dementias. Brain Pathol 22: 99–109.

37. WinslowAR, ChenCW, CorrochanoS, Acevedo-ArozenaA, GordonDE, et al. (2010) Alpha-synuclein impairs macroautophagy: implications for Parkinson's disease. J Cell Biol 190: 1023–1037.

38. Ebrahimi-FakhariD, Cantuti-CastelvetriI, FanZ, RockensteinE, MasliahE, et al. (2011) Distinct roles in vivo for the ubiquitin-proteasome system and the autophagy-lysosomal pathway in the degradation of alpha-synuclein. J Neurosci 31: 14508–14520.

39. YangF, YangYP, MaoCJ, LiuL, ZhengHF, et al. (2013) Crosstalk between the proteasome system and autophagy in the clearance of alpha-synuclein. Acta Pharmacol Sin 34: 674–680.

40. TanikSA, SchultheissCE, Volpicelli-DaleyLA, BrundenKR, LeeVM (2013) Lewy body-like alpha-synuclein aggregates resist degradation and impair macroautophagy. J Biol Chem 288: 15194–15210.

41. ZabrockiP, PellensK, VanhelmontT, VandebroekT, GriffioenG, et al. (2005) Characterization of alpha-synuclein aggregation and synergistic toxicity with protein tau in yeast. FEBS J 272: 1386–1400.

42. SancenonV, LeeSA, PatrickC, GriffithJ, PaulinoA, et al. (2012) Suppression of alpha-synuclein toxicity and vesicle trafficking defects by phosphorylation at S129 in yeast depends on genetic context. Hum Mol Genet 21: 2432–2449.

43. ZabrockiP, BastiaensI, DelayC, BammensT, GhillebertR, et al. (2008) Phosphorylation, lipid raft interaction and traffic of alpha-synuclein in a yeast model for Parkinson. Biochim Biophys Acta 1783: 1767–1780.

44. KaganovichD, KopitoR, FrydmanJ (2008) Misfolded proteins partition between two distinct quality control compartments. Nature 454: 1088–1095.

45. BuchanJR, YoonJH, ParkerR (2011) Stress-specific composition, assembly and kinetics of stress granules in Saccharomyces cerevisiae. J Cell Sci 124: 228–239.

46. PaivaS, VieiraN, NondierI, Haguenauer-TsapisR, CasalM, et al. (2009) Glucose-induced ubiquitylation and endocytosis of the yeast Jen1 transporter: role of lysine 63-linked ubiquitin chains. J Biol Chem 284: 19228–19236.

47. BecuweM, VieiraN, LaraD, Gomes-RezendeJ, Soares-CunhaC, et al. (2012) A molecular switch on an arrestin-like protein relays glucose signaling to transporter endocytosis. J Cell Biol 196: 247–259.

48. CollinsGA, GomezTA, DeshaiesRJ, TanseyWP (2010) Combined chemical and genetic approach to inhibit proteolysis by the proteasome. Yeast 27: 965–974.

49. KlionskyDJ, AbdallaFC, AbeliovichH, AbrahamRT, Acevedo-ArozenaA, et al. (2012) Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8: 445–544.

50. ShintaniT, KlionskyDJ (2004) Cargo proteins facilitate the formation of transport vesicles in the cytoplasm to vacuole targeting pathway. J Biol Chem 279: 29889–29894.

51. KabeyaY, KamadaY, BabaM, TakikawaH, SasakiM, et al. (2005) Atg17 functions in cooperation with Atg1 and Atg13 in yeast autophagy. Mol Biol Cell 16: 2544–2553.

52. StraubM, BredschneiderM, ThummM (1997) AUT3, a serine/threonine kinase gene, is essential for autophagocytosis in Saccharomyces cerevisiae. J Bacteriol 179: 3875–3883.

53. TanidaI, MizushimaN, KiyookaM, OhsumiM, UenoT, et al. (1999) Apg7p/Cvt2p: A novel protein-activating enzyme essential for autophagy. Mol Biol Cell 10: 1367–1379.

54. PaleologouKE, SchmidAW, RospigliosiCC, KimHY, LambertoGR, et al. (2008) Phosphorylation at Ser-129 but not the phosphomimics S129E/D inhibits the fibrillation of alpha-synuclein. J Biol Chem 283: 16895–16905.

55. OueslatiA, FournierM, LashuelHA (2010) Role of post-translational modifications in modulating the structure, function and toxicity of alpha-synuclein: implications for Parkinson's disease pathogenesis and therapies. Prog Brain Res 183: 115–145.

56. GorbatyukOS, LiS, SullivanLF, ChenW, KondrikovaG, et al. (2008) The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease. Proc Natl Acad Sci U S A 105: 763–768.

57. BrownDR (2010) Oligomeric alpha-synuclein and its role in neuronal death. IUBMB Life 62: 334–339.

58. TyedmersJ, MogkA, BukauB (2010) Cellular strategies for controlling protein aggregation. Nat Rev Mol Cell Biol 11: 777–788.

59. OienDB, ShinogleHE, MooreDS, MoskovitzJ (2009) Clearance and phosphorylation of alpha-synuclein are inhibited in methionine sulfoxide reductase a null yeast cells. J Mol Neurosci 39: 323–332.

60. WinnerB, JappelliR, MajiSK, DesplatsPA, BoyerL, et al. (2011) In vivo demonstration that alpha-synuclein oligomers are toxic. Proc Natl Acad Sci U S A 108: 4194–4199.

61. JeddG, RichardsonC, LittR, SegevN (1995) The Ypt1 GTPase is essential for the first two steps of the yeast secretory pathway. J Cell Biol 131: 583–590.

62. McNewJA, SogaardM, LampenNM, MachidaS, YeRR, et al. (1997) Ykt6p, a prenylated SNARE essential for endoplasmic reticulum-Golgi transport. J Biol Chem 272: 17776–17783.

63. CohenM, StutzF, BelgarehN, Haguenauer-TsapisR, DargemontC (2003) Ubp3 requires a cofactor, Bre5, to specifically de-ubiquitinate the COPII protein, Sec23. Nat Cell Biol 5: 661–667.

64. De AntoniA, SchmitzovaJ, TrepteHH, GallwitzD, AlbertS (2002) Significance of GTP hydrolysis in Ypt1p-regulated endoplasmic reticulum to Golgi transport revealed by the analysis of two novel Ypt1-GAPs. J Biol Chem 277: 41023–41031.

65. AntebiA, FinkGR (1992) The yeast Ca(2+)-ATPase homologue, PMR1, is required for normal Golgi function and localizes in a novel Golgi-like distribution. Mol Biol Cell 3: 633–654.

66. PowersET, MorimotoRI, DillinA, KellyJW, BalchWE (2009) Biological and chemical approaches to diseases of proteostasis deficiency. Annu Rev Biochem 78: 959–991.

67. LashuelHA, OverkCR, OueslatiA, MasliahE (2012) The many faces of alpha-synuclein: from structure and toxicity to therapeutic target. Nat Rev Neurosci 14: 38–48.

68. AuluckPK, MeulenerMC, BoniniNM (2005) Mechanisms of suppression of alpha-synuclein seurotoxicity by geldanamycin in Drosophila. J Biol Chem 280: 2873–2878.

69. SpokoiniR, MoldavskiO, NahmiasY, EnglandJL, SchuldinerM, et al. (2012) Confinement to organelle-associated inclusion structures mediates asymmetric inheritance of aggregated protein in budding yeast. Cell Reports 2: 738–747.

70. WeisbergSJ, LyakhovetskyR, WerdigerAC, GitlerAD, SoenY, et al. (2012) Compartmentalization of superoxide dismutase 1 (SOD1G93A) aggregates determines their toxicity. Proc Natl Acad Sci U S A 109: 15811–15816.

71. KomanderD (2009) The emerging complexity of protein ubiquitination. Biochem Soc Trans 37: 937–953.

72. ThompsonLM, AikenCT, KaltenbachLS, AgrawalN, IllesK, et al. (2009) IKK phosphorylates Huntingtin and targets it for degradation by the proteasome and lysosome. J Cell Biol 187: 1083–1099.

73. JeongH, ThenF, MeliaTJJr, MazzulliJR, CuiL, et al. (2009) Acetylation targets mutant huntingtin to autophagosomes for degradation. Cell 137: 60–72.

74. ChauKY, ChingHL, SchapiraAH, CooperJM (2009) Relationship between alpha-synuclein phosphorylation, proteasomal inhibition and cell death: relevance to Parkinson's disease pathogenesis. J Neurochem 110: 1005–1013.

75. MachiyaY, HaraS, ArawakaS, FukushimaS, SatoH, et al. (2010) Phosphorylated alpha-synuclein at Ser-129 is targeted to the proteasome pathway in a ubiquitin-independent manner. J Biol Chem 285: 40732–40744.

76. ProninAN, MorrisAJ, SurguchovA, BenovicJL (2000) Synucleins are a novel class of substrates for G protein-coupled receptor kinases. J Biol Chem 275: 26515–26522.

77. SatoH, KatoT, ArawakaS (2013) The role of Ser129 phosphorylation of alpha-synuclein in neurodegeneration of Parkinson's disease: a review of in vivo models. Rev Neurosci 24: 115–123.

78. MbefoMK, PaleologouKE, BoucharabaA, OueslatiA, SchellH, et al. (2010) Phosphorylation of synucleins by members of the Polo-like kinase family. J Biol Chem 285: 2807–2822.

79. OueslatiA, SchneiderBL, AebischerP, LashuelHA (2013) Polo-like kinase 2 regulates selective autophagic alpha-synuclein clearance and suppresses its toxicity in vivo. Proc Natl Acad Sci U S A 110: E3945–3954.

80. PandeyUB, NieZ, BatleviY, McCrayBA, RitsonGP, et al. (2007) HDAC6 rescues neurodegeneration and provides an essential link between autophagy and the UPS. Nature 447: 859–863.

81. TofarisGK, LayfieldR, SpillantiniMG (2001) Alpha-synuclein metabolism and aggregation is linked to ubiquitin-independent degradation by the proteasome. FEBS Lett 509: 22–26.

82. AncolioK, Alves da CostaC, UedaK, CheclerF (2000) Alpha-synuclein and the Parkinson's disease-related mutant Ala53Thr-alpha-synuclein do not undergo proteasomal degradation in HEK293 and neuronal cells. Neurosci Lett 285: 79–82.

83. LinderssonE, BeedholmR, HojrupP, MoosT, GaiW, et al. (2004) Proteasomal inhibition by alpha-synuclein filaments and oligomers. J Biol Chem 279: 12924–12934.

84. MorozovaKS, PiatkevichKD, GouldTJ, ZhangJ, BewersdorfJ, et al. (2010) Far-red fluorescent protein excitable with red lasers for flow cytometry and superresolution STED nanoscopy. Biophys J 99: L13–15.

85. SuzukiK, KondoC, MorimotoM, OhsumiY (2010) Selective transport of alpha-mannosidase by autophagic pathways: identification of a novel receptor, Atg34p. J Biol Chem 285: 30019–30025.

86. LaporteD, SalinB, Daignan-FornierB, SagotI (2008) Reversible cytoplasmic localization of the proteasome in quiescent yeast cells. J Biol Chem 181: 737–745.

87. AlbertiS, GitlerAD, LindquistS (2007) A suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae. Yeast 24: 913–919.

88. WendlandJ (2003) PCR-based methods facilitate targeted gene manipulations and cloning procedures. Curr Genet 44: 115–123.

89. GraefM, NunnariJ (2011) Mitochondria regulate autophagy by conserved signalling pathways. EMBO J 30: 2101–2114.

90. BassoE, AntasP, MarijanovicZ, GoncalvesS, TenreiroS, et al. (2013) PLK2 modulates alpha-synuclein aggregation in yeast and mammalian cells. Mol Neurobiol 48: 854–862.

91. FariaC, JorgeCD, BorgesN, TenreiroS, OuteiroTF, et al. (2013) Inhibition of formation of alpha-synuclein inclusions by mannosylglycerate in a yeast model of Parkinson's disease. Biochim Biophys Acta 1830: 4065–4072.

92. TaneseN (1997) Small-scale density gradient sedimentation to separate and analyze multiprotein complexes. Methods 12: 224–234.

93. PhairRD, MisteliT (2000) High mobility of proteins in the mammalian cell nucleus. Nature 404: 604–609.

94. SpragueBL, McNallyJG (2005) FRAP analysis of binding: proper and fitting. Trends Cell Biol 15: 84–91.

95. ThomasBJ, RothsteinR (1989) Elevated recombination rates in transcriptionally active DNA. Cell 56: 619–630.

96. SancenonV, LeeSA, PatrickC, GriffithJ, PaulinoA, et al. (2012) Suppression of alpha-synuclein toxicity and vesicle trafficking defects by phosphorylation at S129 in yeast depends on genetic context. Hum Mol Genet 21: 2432–2449.