GTPase Activity and Neuronal Toxicity of Parkinson's Disease–Associated LRRK2 Is Regulated by ArfGAP1
Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common cause of autosomal dominant familial Parkinson's disease (PD) and also contribute to idiopathic PD. LRRK2 encodes a large multi-domain protein with GTPase and kinase activity. Initial data indicates that an intact functional GTPase domain is critically required for LRRK2 kinase activity. PD–associated mutations in LRRK2, including the most common G2019S variant, have variable effects on enzymatic activity but commonly alter neuronal process morphology. The mechanisms underlying the intrinsic and extrinsic regulation of LRRK2 GTPase and kinase activity, and the pathogenic effects of familial mutations, are incompletely understood. Here, we identify a novel functional interaction between LRRK2 and ADP-ribosylation factor GTPase-activating protein 1 (ArfGAP1). LRRK2 and ArfGAP1 interact in vitro in mammalian cells and in vivo in brain, and co-localize in the cytoplasm and at Golgi membranes. PD–associated and functional mutations that alter the GTPase activity of LRRK2 modulate the interaction with ArfGAP1. The GTP hydrolysis activity of LRRK2 is markedly enhanced by ArfGAP1 supporting a role for ArfGAP1 as a GTPase-activating protein for LRRK2. Unexpectedly, ArfGAP1 promotes the kinase activity of LRRK2 suggesting a potential role for GTP hydrolysis in kinase activation. Furthermore, LRRK2 robustly and directly phosphorylates ArfGAP1 in vitro. Silencing of ArfGAP1 expression in primary cortical neurons rescues the neurite shortening phenotype induced by G2019S LRRK2 overexpression, whereas the co-expression of ArfGAP1 and LRRK2 synergistically promotes neurite shortening in a manner dependent upon LRRK2 GTPase activity. Neurite shortening induced by ArfGAP1 overexpression is also attenuated by silencing of LRRK2. Our data reveal a novel role for ArfGAP1 in regulating the GTPase activity and neuronal toxicity of LRRK2; reciprocally, LRRK2 phosphorylates ArfGAP1 and is required for ArfGAP1 neuronal toxicity. ArfGAP1 may represent a promising target for interfering with LRRK2-dependent neurodegeneration in familial and sporadic PD.
Vyšlo v časopise:
GTPase Activity and Neuronal Toxicity of Parkinson's Disease–Associated LRRK2 Is Regulated by ArfGAP1. PLoS Genet 8(2): e32767. doi:10.1371/journal.pgen.1002526
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002526
Souhrn
Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common cause of autosomal dominant familial Parkinson's disease (PD) and also contribute to idiopathic PD. LRRK2 encodes a large multi-domain protein with GTPase and kinase activity. Initial data indicates that an intact functional GTPase domain is critically required for LRRK2 kinase activity. PD–associated mutations in LRRK2, including the most common G2019S variant, have variable effects on enzymatic activity but commonly alter neuronal process morphology. The mechanisms underlying the intrinsic and extrinsic regulation of LRRK2 GTPase and kinase activity, and the pathogenic effects of familial mutations, are incompletely understood. Here, we identify a novel functional interaction between LRRK2 and ADP-ribosylation factor GTPase-activating protein 1 (ArfGAP1). LRRK2 and ArfGAP1 interact in vitro in mammalian cells and in vivo in brain, and co-localize in the cytoplasm and at Golgi membranes. PD–associated and functional mutations that alter the GTPase activity of LRRK2 modulate the interaction with ArfGAP1. The GTP hydrolysis activity of LRRK2 is markedly enhanced by ArfGAP1 supporting a role for ArfGAP1 as a GTPase-activating protein for LRRK2. Unexpectedly, ArfGAP1 promotes the kinase activity of LRRK2 suggesting a potential role for GTP hydrolysis in kinase activation. Furthermore, LRRK2 robustly and directly phosphorylates ArfGAP1 in vitro. Silencing of ArfGAP1 expression in primary cortical neurons rescues the neurite shortening phenotype induced by G2019S LRRK2 overexpression, whereas the co-expression of ArfGAP1 and LRRK2 synergistically promotes neurite shortening in a manner dependent upon LRRK2 GTPase activity. Neurite shortening induced by ArfGAP1 overexpression is also attenuated by silencing of LRRK2. Our data reveal a novel role for ArfGAP1 in regulating the GTPase activity and neuronal toxicity of LRRK2; reciprocally, LRRK2 phosphorylates ArfGAP1 and is required for ArfGAP1 neuronal toxicity. ArfGAP1 may represent a promising target for interfering with LRRK2-dependent neurodegeneration in familial and sporadic PD.
Zdroje
1. BiskupSWestAB 2009 Zeroing in on LRRK2-linked pathogenic mechanisms in Parkinson's disease. Biochim Biophys Acta 1792 625 633
2. GasserT 2009 Mendelian forms of Parkinson's disease. Biochim Biophys Acta 1792 587 596
3. HealyDGFalchiMO'SullivanSSBonifatiVDurrA 2008 Phenotype, genotype, and worldwide genetic penetrance of LRRK2-associated Parkinson's disease: a case-control study. Lancet Neurol 7 583 590
4. LesageSDurrATazirMLohmannELeuteneggerAL 2006 LRRK2 G2019S as a cause of Parkinson's disease in North African Arabs. N Engl J Med 354 422 423
5. OzeliusLJSenthilGSaunders-PullmanROhmannEDeligtischA 2006 LRRK2 G2019S as a cause of Parkinson's disease in Ashkenazi Jews. N Engl J Med 354 424 425
6. GilksWPAbou-SleimanPMGandhiSJainSSingletonA 2005 A common LRRK2 mutation in idiopathic Parkinson's disease. Lancet 365 415 416
7. MarinIvan EgmondWNvan HaastertPJ 2008 The Roco protein family: a functional perspective. Faseb J 22 3103 3110
8. GloecknerCJKinklNSchumacherABraunRJO'NeillE 2006 The Parkinson disease causing LRRK2 mutation I2020T is associated with increased kinase activity. Hum Mol Genet 15 223 232
9. GreggioEJainSKingsburyABandopadhyayRLewisP 2006 Kinase activity is required for the toxic effects of mutant LRRK2/dardarin. Neurobiol Dis 23 329 341
10. WestABMooreDJBiskupSBugayenkoASmithWW 2005 Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity. Proc Natl Acad Sci U S A 102 16842 16847
11. ImaiYGehrkeSWangHQTakahashiRHasegawaK 2008 Phosphorylation of 4E-BP by LRRK2 affects the maintenance of dopaminergic neurons in Drosophila. Embo J 27 2432 2443
12. JaleelMNicholsRJDeakMCampbellDGGillardonF 2007 LRRK2 phosphorylates moesin at threonine-558: characterization of how Parkinson's disease mutants affect kinase activity. Biochem J 405 307 317
13. GillardonF 2009 Leucine-rich repeat kinase 2 phosphorylates brain tubulin-beta isoforms and modulates microtubule stability–a point of convergence in parkinsonian neurodegeneration? J Neurochem 110 1514 1522
14. KanaoTVenderovaKParkDSUntermanTLuB 2010 Activation of FoxO by LRRK2 induces expression of proapoptotic proteins and alters survival of postmitotic dopaminergic neuron in Drosophila. Hum Mol Genet 19 3747 3758
15. KumarAGreggioEBeilinaAKaganovichAChanD 2010 The Parkinson's disease associated LRRK2 exhibits weaker in vitro phosphorylation of 4E-BP compared to autophosphorylation. PLoS ONE 5 e8730 doi:10.1371/journal.pone.0008730
16. ItoGOkaiTFujinoGTakedaKIchijoH 2007 GTP binding is essential to the protein kinase activity of LRRK2, a causative gene product for familial Parkinson's disease. Biochemistry 46 1380 1388
17. LewisPAGreggioEBeilinaAJainSBakerA 2007 The R1441C mutation of LRRK2 disrupts GTP hydrolysis. Biochem Biophys Res Commun 357 668 671
18. LiXTanYCPouloseSOlanowCWHuangXY 2007 Leucine-rich repeat kinase 2 (LRRK2)/PARK8 possesses GTPase activity that is altered in familial Parkinson's disease R1441C/G mutants. J Neurochem 103 238 247
19. SmithWWPeiZJiangHDawsonVLDawsonTM 2006 Kinase activity of mutant LRRK2 mediates neuronal toxicity. Nat Neurosci 9 1231 1233
20. WestABMooreDJChoiCAndrabiSALiX 2007 Parkinson's disease-associated mutations in LRRK2 link enhanced GTP-binding and kinase activities to neuronal toxicity. Hum Mol Genet 16 223 232
21. TaymansJMVancraenenbroeckROllikainenPBeilinaALobbestaelE 2011 LRRK2 kinase activity is dependent on LRRK2 GTP binding capacity but independent of LRRK2 GTP binding. PLoS ONE 6 e23207 doi:10.1371/journal.pone.0023207
22. GloecknerCJBoldtKvon ZweydorfFHelmSWiesentL 2010 Phosphopeptide analysis reveals two discrete clusters of phosphorylation in the N-terminus and the Roc domain of the Parkinson-disease associated protein kinase LRRK2. J Proteome Res 9 1738 1745
23. GreggioETaymansJMZhenEYRyderJVancraenenbroeckR 2009 The Parkinson's disease kinase LRRK2 autophosphorylates its GTPase domain at multiple sites. Biochem Biophys Res Commun 389 449 454
24. KamikawajiSItoGIwatsuboT 2009 Identification of the autophosphorylation sites of LRRK2. Biochemistry 48 10963 10975
25. WebberPJSmithADSenSRenfrowMBMobleyJA 2011 Autophosphorylation in the leucine-rich repeat kinase 2 (LRRK2) GTPase domain modifies kinase and GTP-binding activities. J Mol Biol 412 94 110
26. HaebigKGloecknerCJMirallesMGGillardonFSchulteC 2010 ARHGEF7 (Beta-PIX) acts as guanine nucleotide exchange factor for leucine-rich repeat kinase 2. PLoS ONE 5 e13762 doi:10.1371/journal.pone.0013762
27. XiongYCoombesCEKilaruALiXGitlerAD 2010 GTPase activity plays a key role in the pathobiology of LRRK2. PLoS Genet 6 e1000902 doi:10.1371/journal.pgen.1000902
28. DanielsVVancraenenbroeckRLawBMGreggioELobbestaelE 2011 Insight into the mode of action of the LRRK2 Y1699C pathogenic mutant. J Neurochem 116 304 315
29. IaccarinoCCrosioCVitaleCSannaGCarriMT 2007 Apoptotic mechanisms in mutant LRRK2-mediated cell death. Hum Mol Genet 16 1319 1326
30. SmithWWPeiZJiangHMooreDJLiangY 2005 Leucine-rich repeat kinase 2 (LRRK2) interacts with parkin, and mutant LRRK2 induces neuronal degeneration. Proc Natl Acad Sci U S A 102 18676 18681
31. HoCCRideoutHJRibeETroyCMDauerWT 2009 The Parkinson disease protein leucine-rich repeat kinase 2 transduces death signals via Fas-associated protein with death domain and caspase-8 in a cellular model of neurodegeneration. J Neurosci 29 1011 1016
32. DusonchetJKochubeyOStafaKYoungSMJrZuffereyR 2011 A Rat Model of Progressive Nigral Neurodegeneration Induced by the Parkinson's Disease-Associated G2019S Mutation in LRRK2. J Neurosci 31 907 912
33. LeeBDShinJHVanKampenJPetrucelliLWestAB 2010 Inhibitors of leucine-rich repeat kinase-2 protect against models of Parkinson's disease. Nat Med 16 998 1000
34. LiYLiuWOoTFWangLTangY 2009 Mutant LRRK2(R1441G) BAC transgenic mice recapitulate cardinal features of Parkinson's disease. Nat Neurosci 12 826 828
35. RamonetDDaherJPLinBMStafaKKimJ 2011 Dopaminergic neuronal loss, reduced neurite complexity and autophagic abnormalities in transgenic mice expressing G2019S mutant LRRK2. PLoS ONE 6 e18568 doi:10.1371/journal.pone.0018568
36. MacLeodDDowmanJHammondRLeeteTInoueK 2006 The familial Parkinsonism gene LRRK2 regulates neurite process morphology. Neuron 52 587 593
37. PloweyEDCherraSJ3rdLiuYJChuCT 2008 Role of autophagy in G2019S-LRRK2-associated neurite shortening in differentiated SH-SY5Y cells. J Neurochem 105 1048 1056
38. ParisiadouLXieCChoHJLinXGuXL 2009 Phosphorylation of ezrin/radixin/moesin proteins by LRRK2 promotes the rearrangement of actin cytoskeleton in neuronal morphogenesis. J Neurosci 29 13971 13980
39. CukiermanEHuberIRotmanMCasselD 1995 The ARF1 GTPase-activating protein: zinc finger motif and Golgi complex localization. Science 270 1999 2002
40. LiuWDudenRPhairRDLippincott-SchwartzJ 2005 ArfGAP1 dynamics and its role in COPI coat assembly on Golgi membranes of living cells. J Cell Biol 168 1053 1063
41. DonaldsonJGCasselDKahnRAKlausnerRD 1992 ADP-ribosylation factor, a small GTP-binding protein, is required for binding of the coatomer protein beta-COP to Golgi membranes. Proc Natl Acad Sci U S A 89 6408 6412
42. TaylorTCKahnRAMelanconP 1992 Two distinct members of the ADP-ribosylation factor family of GTP-binding proteins regulate cell-free intra-Golgi transport. Cell 70 69 79
43. DonaldsonJGJacksonCL 2011 ARF family G proteins and their regulators: roles in membrane transport, development and disease. Nat Rev Mol Cell Biol 12 362 375
44. BiskupSMooreDJCelsiFHigashiSWestAB 2006 Localization of LRRK2 to membranous and vesicular structures in mammalian brain. Ann Neurol 60 557 569
45. HatanoTKuboSImaiSMaedaMIshikawaK 2007 Leucine-rich repeat kinase 2 associates with lipid rafts. Hum Mol Genet 16 678 690
46. Alegre-AbarrateguiJChristianHLufinoMMMutihacRVendaLL 2009 LRRK2 regulates autophagic activity and localizes to specific membrane microdomains in a novel human genomic reporter cellular model. Hum Mol Genet 18 4022 4034
47. ParnisARawetMRegevLBarkanBRotmanM 2006 Golgi localization determinants in ArfGAP1 and in new tissue-specific ArfGAP1 isoforms. J Biol Chem 281 3785 3792
48. KanaoTVenderovaKParkDSUntermanTLuB 2010 Activation of FoxO by LRRK2 induces expression of proapoptotic proteins and alters survival of postmitotic dopaminergic neuron in Drosophila. Hum Mol Genet
49. WangLXieCGreggioEParisiadouLShimH 2008 The chaperone activity of heat shock protein 90 is critical for maintaining the stability of leucine-rich repeat kinase 2. J Neurosci 28 3384 3391
50. BiskupSMooreDJReaALorenz-DeperieuxBCoombesCE 2007 Dynamic and redundant regulation of LRRK2 and LRRK1 expression. BMC Neurosci 8 102
51. HigashiSBiskupSWestABTrinkausDDawsonVL 2007 Localization of Parkinson's disease-associated LRRK2 in normal and pathological human brain. Brain Res 1155 208 219
52. HigashiSMooreDJColebrookeREBiskupSDawsonVL 2007 Expression and localization of Parkinson's disease-associated leucine-rich repeat kinase 2 in the mouse brain. J Neurochem 100 368 381
53. McMahonHTMillsIG 2004 COP and clathrin-coated vesicle budding: different pathways, common approaches. Curr Opin Cell Biol 16 379 391
54. LeeSYYangJSHongWPremontRTHsuVW 2005 ARFGAP1 plays a central role in coupling COPI cargo sorting with vesicle formation. J Cell Biol 168 281 290
55. YangJSLeeSYGaoMBourgoinSRandazzoPA 2002 ARFGAP1 promotes the formation of COPI vesicles, suggesting function as a component of the coat. J Cell Biol 159 69 78
56. Lippincott-SchwartzJLiuW 2006 Insights into COPI coat assembly and function in living cells. Trends Cell Biol 16 e1 4
57. BeckRAdolfFWeimerCBrueggerBWielandFT 2009 ArfGAP1 activity and COPI vesicle biogenesis. Traffic 10 307 315
58. BaiMGadHTuracchioGCocucciEYangJS 2011 ARFGAP1 promotes AP-2-dependent endocytosis. Nat Cell Biol 13 559 567
59. MarinI 2006 The Parkinson disease gene LRRK2: evolutionary and structural insights. Mol Biol Evol 23 2423 2433
60. DengJLewisPAGreggioESluchEBeilinaA 2008 Structure of the ROC domain from the Parkinson's disease-associated leucine-rich repeat kinase 2 reveals a dimeric GTPase. Proc Natl Acad Sci U S A 105 1499 1504
61. LiXMooreDJXiongYDawsonTMDawsonVL 2010 Reevaluation of phosphorylation sites in the Parkinson disease-associated leucine-rich repeat kinase 2. J Biol Chem 285 29569 29576
62. SenSWebberPJWestAB 2009 Dependence of leucine-rich repeat kinase 2 (LRRK2) kinase activity on dimerization. J Biol Chem 284 36346 36356
63. BergerZSmithKALavoieMJ 2010 Membrane localization of LRRK2 is associated with increased formation of the highly active LRRK2 dimer and changes in its phosphorylation. Biochemistry 49 5511 5523
64. LinXParisiadouLGuXLWangLShimH 2009 Leucine-rich repeat kinase 2 regulates the progression of neuropathology induced by Parkinson's-disease-related mutant alpha-synuclein. Neuron 64 807 827
65. ShinNJeongHKwonJHeoHYKwonJJ 2008 LRRK2 regulates synaptic vesicle endocytosis. Exp Cell Res 314 2055 2065
66. PiccoliGCondliffeSBBauerMGiesertFBoldtK 2011 LRRK2 controls synaptic vesicle storage and mobilization within the recycling pool. J Neurosci 31 2225 2237
67. MeixnerABoldtKVan TroysMAskenaziMGloecknerCJ 2011 A QUICK screen for Lrrk2 interaction partners–leucine-rich repeat kinase 2 is involved in actin cytoskeleton dynamics. Mol Cell Proteomics 10 M110 001172
68. BeemillerPHoppeADSwansonJA 2006 A phosphatidylinositol-3-kinase-dependent signal transition regulates ARF1 and ARF6 during Fcgamma receptor-mediated phagocytosis. PLoS Biol 4 e162 doi:10.1371/journal.pbio.0040162
69. MooreDJWestABDikemanDADawsonVLDawsonTM 2008 Parkin mediates the degradation-independent ubiquitination of Hsp70. J Neurochem 105 1806 1819
70. StrackRLStronginDEBhattacharyyaDTaoWBermanA 2008 A noncytotoxic DsRed variant for whole-cell labeling. Nat Methods 5 955 957
71. SarbassovDDGuertinDAAliSMSabatiniDM 2005 Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307 1098 1101
72. Luthi-CarterRTaylorDMPallosJLambertEAmoreA 2010 SIRT2 inhibition achieves neuroprotection by decreasing sterol biosynthesis. Proc Natl Acad Sci U S A 107 7927 7932
73. HerzigMCKollyCPersohnETheilDSchweizerT 2011 LRRK2 protein levels are determined by kinase function and are crucial for kidney and lung homeostasis in mice. Hum Mol Genet 20 4209 4223
74. HallettPJCollinsTLStandaertDGDunahAW 2008 Biochemical fractionation of brain tissue for studies of receptor distribution and trafficking. Curr Protoc Neurosci Chapter 1 Unit 1 16
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2012 Číslo 2
- Je „freeze-all“ pro všechny? Odborníci na fertilitu diskutovali na virtuálním summitu
- Gynekologové a odborníci na reprodukční medicínu se sejdou na prvním virtuálním summitu
Najčítanejšie v tomto čísle
- Gene Expression and Stress Response Mediated by the Epigenetic Regulation of a Transposable Element Small RNA
- Contrasting Properties of Gene-Specific Regulatory, Coding, and Copy Number Mutations in : Frequency, Effects, and Dominance
- Homeobox Genes Critically Regulate Embryo Implantation by Controlling Paracrine Signaling between Uterine Stroma and Epithelium
- Nondisjunction of a Single Chromosome Leads to Breakage and Activation of DNA Damage Checkpoint in G2