BACKGROUND: Autosomal-dominant mutations in the Park8 gene encoding Leucine-rich repeat kinase 2 (LRRK2) have been identified to cause up to 40% of the genetic forms of Parkinson's disease. However, the function and molecular pathways regulated by LRRK2 are largely unknown. It has been shown that LRRK2 serves as a scaffold during activation of WNT/β-catenin signaling via its interaction with the β-catenin destruction complex, DVL1-3 and LRP6. In this study, we examine whether LRRK2 also interacts with signaling components of the WNT/Planar Cell Polarity (WNT/PCP) pathway, which controls the maturation of substantia nigra dopaminergic neurons, the main cell type lost in Parkinson's disease patients. METHODS: Co-immunoprecipitation and tandem mass spectrometry was performed in a mouse substantia nigra cell line (SN4741) and human HEK293T cell line in order to identify novel LRRK2 binding partners. Inhibition of the WNT/β-catenin reporter, TOPFlash, was used as a read-out of WNT/PCP pathway activation. The capacity of LRRK2 to regulate WNT/PCP signaling in vivo was tested in Xenopus laevis' early development. RESULTS: Our proteomic analysis identified that LRRK2 interacts with proteins involved in WNT/PCP signaling such as the PDZ domain-containing protein GIPC1 and Integrin-linked kinase (ILK) in dopaminergic cells in vitro and in the mouse ventral midbrain in vivo. Moreover, co-immunoprecipitation analysis revealed that LRRK2 binds to two core components of the WNT/PCP signaling pathway, PRICKLE1 and CELSR1, as well as to FLOTILLIN-2 and CULLIN-3, which regulate WNT secretion and inhibit WNT/β-catenin signaling, respectively. We also found that PRICKLE1 and LRRK2 localize in signalosomes and act as dual regulators of WNT/PCP and β-catenin signaling. Accordingly, analysis of the function of LRRK2 in vivo, in X. laevis revelaed that LRKK2 not only inhibits WNT/β-catenin pathway, but induces a classical WNT/PCP phenotype in vivo. CONCLUSIONS: Our study shows for the first time that LRRK2 activates the WNT/PCP signaling pathway through its interaction to multiple WNT/PCP components. We suggest that LRRK2 regulates the balance between WNT/β-catenin and WNT/PCP signaling, depending on the binding partners. Since this balance is crucial for homeostasis of midbrain dopaminergic neurons, we hypothesize that its alteration may contribute to the pathophysiology of Parkinson's disease.

Central European Institute for Technology Masaryk University Kamenice 5 625 00 Brno Czech Republic

Current address Science for Life Laboratory Department of Biophysics and Biochemistry Stockholm University 171 65 Stockholm Sweden

Department of Experimental Biology Faculty of Science Masaryk University 625 00 Brno Czech Republic

Laboratory of Molecular Neurobiology Department of Medical Biochemistry and Biophysics Karolinska Institutet 17177 Stockholm Sweden

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Lin MK, Farrer MJ. Genetics and genomics of Parkinson's disease. Genome Med. 2014;6:48. doi: 10.1186/gm566. PubMed DOI PMC

Dachsel JC, Nishioka K, Vilarino-Guell C, Lincoln SJ, Soto-Ortolaza AI, Kachergus J, Hinkle KM, Heckman MG, Jasinska-Myga B, Taylor JP, et al. Heterodimerization of Lrrk1-Lrrk2: implications for LRRK2-associated Parkinson disease. Mech Ageing Dev. 2010;131:210–214. doi: 10.1016/j.mad.2010.01.009. PubMed DOI PMC

Klein C, Westenberger A. Genetics of Parkinson's disease. Cold Spring Harb Perspect Med. 2012;2:a008888. doi: 10.1101/cshperspect.a008888. PubMed DOI PMC

Wood-Kaczmar A, Gandhi S, Wood NW. Understanding the molecular causes of Parkinson's disease. Trends Mol Med. 2006;12:521–528. doi: 10.1016/j.molmed.2006.09.007. PubMed DOI

Kumaran R, Cookson MR. Pathways to parkinsonism Redux: convergent pathobiological mechanisms in genetics of Parkinson's disease. Hum Mol Genet. 2015;24:R32–R44. doi: 10.1093/hmg/ddv236. PubMed DOI PMC

Funayama M, Hasegawa K, Kowa H, Saito M, Tsuji S, Obata F. A new locus for Parkinson's disease (PARK8) maps to chromosome 12p11.2-q13.1. Ann Neurol. 2002;51:296–301. doi: 10.1002/ana.10113. PubMed DOI

Zimprich A, Biskup S, Leitner P, Lichtner P, Farrer M, Lincoln S, Kachergus J, Hulihan M, Uitti RJ, Calne DB, et al. Mutations in LRRK2 cause autosomal-dominant parkinsonism with pleomorphic pathology. Neuron. 2004;44:601–607. doi: 10.1016/j.neuron.2004.11.005. PubMed DOI

Marin I, van Egmond WN, van Haastert PJ. The Roco protein family: a functional perspective. FASEB J. 2008;22:3103–3110. doi: 10.1096/fj.08-111310. PubMed DOI

Angers S, Moon RT. Proximal events in Wnt signal transduction. Nat Rev Mol Cell Biol. 2009;10:468–477. doi: 10.1038/nrn2674. PubMed DOI

Salinas PC. Wnt signaling in the vertebrate central nervous system: from axon guidance to synaptic function. Cold Spring Harb Perspect Biol. 2012;4(2):a008003. doi: 10.1101/cshperspect.a008003. PubMed DOI PMC

Niehrs C. The complex world of WNT receptor signalling. Nat Rev Mol Cell Biol. 2012;13:767–779. doi: 10.1038/nrm3470. PubMed DOI

Sato A, Yamamoto H, Sakane H, Koyama H, Kikuchi A. Wnt5a regulates distinct signalling pathways by binding to Frizzled2. EMBO J. 2010;29:41–54. doi: 10.1038/emboj.2009.322. PubMed DOI PMC

Andersson ER, Prakash N, Cajanek L, Minina E, Bryja V, Bryjova L, Yamaguchi TP, Hall AC, Wurst W, Arenas E. Wnt5a regulates ventral midbrain morphogenesis and the development of A9-A10 dopaminergic cells in vivo. PLoS One. 2008;3:e3517. doi: 10.1371/journal.pone.0003517. PubMed DOI PMC

Andersson ER, Salto C, Villaescusa JC, Cajanek L, Yang S, Bryjova L, Nagy VSJ, II, Ramirez C, Bryja V, Arenas E. Wnt5a cooperates with canonical Wnts to generate midbrain dopaminergic neurons in vivo and in stem cells. Proc Natl Acad Sci U S A. 2013;110:E602–E610. doi: 10.1073/pnas.1208524110. PubMed DOI PMC

Inestrosa NC, Arenas E. Emerging roles of Wnts in the adult nervous system. Nat Rev Neurosci. 2010;11:77–86. doi: 10.1038/nrn2755. PubMed DOI

Arenas E. Wnt signaling in midbrain dopaminergic neuron development and regenerative medicine for Parkinson's disease. J Mol Cell Biol. 2014;6:42–53. doi: 10.1093/jmcb/mju001. PubMed DOI

Xiong H, Wang D, Chen L, Choo YS, Ma H, Tang C, Xia K, Jiang W, Ronai Z, Zhuang X, Zhang Z. Parkin, PINK1, and DJ-1 form a ubiquitin E3 ligase complex promoting unfolded protein degradation. J Clin Invest. 2009;119:650–660. doi: 10.1172/JCI37617. PubMed DOI PMC

Rawal N, Corti O, Sacchetti P, Ardilla-Osorio H, Sehat B, Brice A, Arenas E. Parkin protects dopaminergic neurons from excessive Wnt/beta-catenin signaling. Biochem Biophys Res Commun. 2009;388:473–478. doi: 10.1016/j.bbrc.2009.07.014. PubMed DOI

Sancho RM, Law BM, Harvey K. Mutations in the LRRK2 roc-COR tandem domain link Parkinson's disease to Wnt signalling pathways. Hum Mol Genet. 2009;18:3955–3968. doi: 10.1093/hmg/ddp337. PubMed DOI PMC

Berwick DC, Harvey K. LRRK2 functions as a Wnt signaling scaffold, bridging cytosolic proteins and membrane-localized LRP6. Hum Mol Genet. 2012;21:4966–4979. doi: 10.1093/hmg/dds342. PubMed DOI PMC

Giese AP, Ezan J, Wang L, Lasvaux L, Lembo F, Mazzocco C, Richard E, Reboul J, Borg JP, Kelley MW, et al. Gipc1 has a dual role in Vangl2 trafficking and hair bundle integrity in the inner ear. Development. 2012;139:3775–3785. doi: 10.1242/dev.074229. PubMed DOI PMC

Perez VA, Ali Z, Alastalo TP, Ikeno F, Sawada H, Lai YJ, Kleisli T, Spiekerkoetter E, Qu X, Rubinos LH, et al. BMP promotes motility and represses growth of smooth muscle cells by activation of tandem Wnt pathways. J Cell Biol. 2011;192:171–188. doi: 10.1083/jcb.201008060. PubMed DOI PMC

Rudkouskaya A, Welch I, Dagnino L. ILK modulates epithelial polarity and matrix formation in hair follicles. Mol Biol Cell. 2014;25:620–632. doi: 10.1091/mbc.E13-08-0499. PubMed DOI PMC

Tao H, Suzuki M, Kiyonari H, Abe T, Sasaoka T, Ueno N. Mouse prickle1, the homolog of a PCP gene, is essential for epiblast apical-basal polarity. Proc Natl Acad Sci U S A. 2009;106:14426–14431. doi: 10.1073/pnas.0901332106. PubMed DOI PMC

Liu C, Lin C, Gao C, May-Simera H, Swaroop A, Li T. Null and hypomorph Prickle1 alleles in mice phenocopy human Robinow syndrome and disrupt signaling downstream of Wnt5a. Biology open. 2014;3:861–870. doi: 10.1242/bio.20148375. PubMed DOI PMC

Curtin JA, Quint E, Tsipouri V, Arkell RM, Cattanach B, Copp AJ, Henderson DJ, Spurr N, Stanier P, Fisher EM, et al. Mutation of Celsr1 disrupts planar polarity of inner ear hair cells and causes severe neural tube defects in the mouse. Curr Biol. 2003;13:1129–1133. doi: 10.1016/S0960-9822(03)00374-9. PubMed DOI

Morgan R, El-Kadi AM, Theokli C. Flamingo, a cadherin-type receptor involved in the drosophila planar polarity pathway, can block signaling via the canonical wnt pathway in Xenopus laevis. Int J Dev Biol. 2003;47:245–252. PubMed

Son JH, Chun HS, Joh TH, Cho S, Conti B, Lee JW. Neuroprotection and neuronal differentiation studies using substantia nigra dopaminergic cells derived from transgenic mouse embryos. J Neurosci. 1999;19:10–20. PubMed PMC

Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science (New York, NY) 2014;343:84–87. doi: 10.1126/science.1247005. PubMed DOI PMC

Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 2015;517:583–588. doi: 10.1038/nature14136. PubMed DOI PMC

Nieuwkoop P, Faber J. Normal table of Xenopus laevis (Daudin): a systematical and chronological survey of the development from the fertilized egg till the end of metamorphosis. 1994.

Migheli R, Del Giudice MG, Spissu Y, Sanna G, Xiong Y, Dawson TM, Dawson VL, Galioto M, Rocchitta G, Biosa A, et al. LRRK2 affects vesicle trafficking, neurotransmitter extracellular level and membrane receptor localization. PLoS One. 2013;8:e77198. doi: 10.1371/journal.pone.0077198. PubMed DOI PMC

Stenman JM, Rajagopal J, Carroll TJ, Ishibashi M, McMahon J, McMahon AP. Canonical Wnt signaling regulates organ-specific assembly and differentiation of CNS vasculature. Science (New York, NY) 2008;322:1247–1250. doi: 10.1126/science.1164594. PubMed DOI

UniProt: a hub for protein information. Nucleic Acids Res 2015, 43:D204-D212. PubMed PMC

Hernandez-Valladares M, Aasebo E, Selheim F, Berven FS, Bruserud O. Selecting sample preparation workflows for mass spectrometry-based proteomic and Phosphoproteomic analysis of patient samples with acute myeloid leukemia. Proteomes. 2016;4 PubMed PMC

Aasebo E, Mjaavatten O, Vaudel M, Farag Y, Selheim F, Berven F, Bruserud O, Hernandez-Valladares M. Freezing effects on the acute myeloid leukemia cell proteome and phosphoproteome revealed using optimal quantitative workflows. J Proteomics. 2016;145:214–225. doi: 10.1016/j.jprot.2016.03.049. PubMed DOI

Terry DE, Umstot E, Desiderio DM. Optimized sample-processing time and peptide recovery for the mass spectrometric analysis of protein digests. J Am Soc Mass Spectrom. 2004;15:784–794. doi: 10.1016/j.jasms.2004.02.005. PubMed DOI

Novak A, Hsu SC, Leung-Hagesteijn C, Radeva G, Papkoff J, Montesano R, Roskelley C, Grosschedl R, Dedhar S. Cell adhesion and the integrin-linked kinase regulate the LEF-1 and beta-catenin signaling pathways. Proc Natl Acad Sci U S A. 1998;95:4374–4379. doi: 10.1073/pnas.95.8.4374. PubMed DOI PMC

Rallis C, Pinchin SM, Ish-Horowicz D. Cell-autonomous integrin control of Wnt and notch signalling during somitogenesis. Development. 2010;137:3591–3601. doi: 10.1242/dev.050070. PubMed DOI

Hsu CH, Chan D, Wolozin B. LRRK2 and the stress response: interaction with MKKs and JNK-interacting proteins. Neurodegener Dis. 2010;7:68–75. doi: 10.1159/000285509. PubMed DOI PMC

Habig K, Walter M, Poths S, Riess O, Bonin M. RNA interference of LRRK2-microarray expression analysis of a Parkinson's disease key player. Neurogenetics. 2008;9:83–94. doi: 10.1007/s10048-007-0114-0. PubMed DOI

Angers S, Thorpe CJ, Biechele TL, Goldenberg SJ, Zheng N, MacCoss MJ, Moon RT. The KLHL12-Cullin-3 ubiquitin ligase negatively regulates the Wnt-beta-catenin pathway by targeting Dishevelled for degradation. Nat Cell Biol. 2006;8:348–357. doi: 10.1038/ncb1381. PubMed DOI

Smalley MJ, Signoret N, Robertson D, Tilley A, Hann A, Ewan K, Ding Y, Paterson H, Dale TC. Dishevelled (Dvl-2) activates canonical Wnt signalling in the absence of cytoplasmic puncta. J Cell Sci. 2005;118:5279–5289. doi: 10.1242/jcs.02647. PubMed DOI

Axelrod JD, Miller JR, Shulman JM, Moon RT, Perrimon N. Differential recruitment of Dishevelled provides signaling specificity in the planar cell polarity and wingless signaling pathways. Genes Dev. 1998;12:2610–2622. doi: 10.1101/gad.12.16.2610. PubMed DOI PMC

Cliffe A, Hamada F, Bienz M. A role of Dishevelled in relocating Axin to the plasma membrane during wingless signaling. Current biology : CB. 2003;13:960–966. doi: 10.1016/S0960-9822(03)00370-1. PubMed DOI

Cajanek L, Ganji RS, Henriques-Oliveira C, Theofilopoulos S, Konik P, Bryja V, Arenas E. Tiam1 regulates the Wnt/Dvl/Rac1 signaling pathway and the differentiation of midbrain dopaminergic neurons. Mol Cell Biol. 2013;33:59–70. doi: 10.1128/MCB.00745-12. PubMed DOI PMC

Gomez-Suaga P, Rivero-Rios P, Fdez E, Blanca Ramirez M, Ferrer I, Aiastui A, Lopez De Munain A, Hilfiker S, et al. Hum Mol Genet. 2014;23:6779–6796. doi: 10.1093/hmg/ddu395. PubMed DOI

MacLeod DA, Rhinn H, Kuwahara T, Zolin A, Di Paolo G, McCabe BD, Marder KS, Honig LS, Clark LN, Small SA, Abeliovich A. RAB7L1 interacts with LRRK2 to modify intraneuronal protein sorting and Parkinson's disease risk. Neuron. 2013;77:425–439. doi: 10.1016/j.neuron.2012.11.033. PubMed DOI PMC

Dodson MW, Zhang T, Jiang C, Chen S, Guo M. Roles of the drosophila LRRK2 homolog in Rab7-dependent lysosomal positioning. Hum Mol Genet. 2012;21:1350–1363. doi: 10.1093/hmg/ddr573. PubMed DOI PMC

Shin N, Jeong H, Kwon J, Heo HY, Kwon JJ, Yun HJ, Kim CH, Han BS, Tong Y, Shen J, et al. LRRK2 regulates synaptic vesicle endocytosis. Exp Cell Res. 2008;314:2055–2065. doi: 10.1016/j.yexcr.2008.02.015. PubMed DOI

Gonzalez-Sancho JM, Greer YE, Abrahams CL, Takigawa Y, Baljinnyam B, Lee KH, Lee KS, Rubin JS, Brown AM. Functional consequences of Wnt-induced dishevelled 2 phosphorylation in canonical and noncanonical Wnt signaling. J Biol Chem. 2013;288:9428–9437. doi: 10.1074/jbc.M112.448480. PubMed DOI PMC

Gao C, Chen YG. Dishevelled: the hub of Wnt signaling. Cell Signal. 2010;22:717–727. doi: 10.1016/j.cellsig.2009.11.021. PubMed DOI

Lee YN, Gao Y, Wang HY. Differential mediation of the Wnt canonical pathway by mammalian Dishevelleds-1, −2, and −3. Cell Signal. 2008;20:443–452. doi: 10.1016/j.cellsig.2007.11.005. PubMed DOI PMC

Chan DW, Chan CY, Yam JW, Ching YP, Ng IO. Prickle-1 negatively regulates Wnt/beta-catenin pathway by promoting Dishevelled ubiquitination/degradation in liver cancer. Gastroenterology. 2006;131:1218–1227. doi: 10.1053/j.gastro.2006.07.020. PubMed DOI

Tree DR, Shulman JM, Rousset R, Scott MP, Gubb D, Axelrod JD. Prickle mediates feedback amplification to generate asymmetric planar cell polarity signaling. Cell. 2002;109:371–381. doi: 10.1016/S0092-8674(02)00715-8. PubMed DOI

Lin YY, Gubb D. Molecular dissection of drosophila prickle isoforms distinguishes their essential and overlapping roles in planar cell polarity. Dev Biol. 2009;325:386–399. doi: 10.1016/j.ydbio.2008.10.042. PubMed DOI

Sweede M, Ankem G, Chutvirasakul B, Azurmendi HF, Chbeir S, Watkins J, Helm RF, Finkielstein CV, Capelluto DG. Structural and membrane binding properties of the prickle PET domain. Biochemistry. 2008;47:13524–13536. doi: 10.1021/bi801037h. PubMed DOI

Cookson MR. LRRK2 pathways leading to Neurodegeneration. Curr Neurol Neurosci Rep. 2015;15:42. doi: 10.1007/s11910-015-0564-y. PubMed DOI PMC

Skibinski G, Nakamura K, Cookson MR, Finkbeiner S. Mutant LRRK2 toxicity in neurons depends on LRRK2 levels and synuclein but not kinase activity or inclusion bodies. J Neurosci. 2014;34:418–433. doi: 10.1523/JNEUROSCI.2712-13.2014. PubMed DOI PMC

Reynolds A, Doggett EA, Riddle SM, Lebakken CS, Nichols RJ. LRRK2 kinase activity and biology are not uniformly predicted by its autophosphorylation and cellular phosphorylation site status. Front Mol Neurosci. 2014;7:54. doi: 10.3389/fnmol.2014.00054. PubMed DOI PMC

Volta M, Cataldi S, Beccano-Kelly D, Munsie L, Tatarnikov I, Chou P, Bergeron S, Mitchell E, Lim R, Khinda J, et al. Chronic and acute LRRK2 silencing has no long-term behavioral effects, whereas wild-type and mutant LRRK2 overexpression induce motor and cognitive deficits and altered regulation of dopamine release. Parkinsonism Relat Disord. 2015;21:1156–1163. doi: 10.1016/j.parkreldis.2015.07.025. PubMed DOI

Hikasa H, Sokol SY. Wnt signaling in vertebrate axis specification. Cold Spring Harb Perspect Biol. 2013;5:a007955. doi: 10.1101/cshperspect.a007955. PubMed DOI PMC

Wallingford JB, Rowning BA, Vogeli KM, Rothbacher U, Fraser SE, Harland RM. Dishevelled controls cell polarity during Xenopus gastrulation. Nature. 2000;405:81–85. doi: 10.1038/35011077. PubMed DOI

Gao B. Wnt regulation of planar cell polarity (PCP) Curr Top Dev Biol. 2012;101:263–295. doi: 10.1016/B978-0-12-394592-1.00008-9. PubMed DOI

Carreira-Barbosa F, Kajita M, Morel V, Wada H, Okamoto H, Martinez Arias A, Fujita Y, Wilson SW, Tada M. Flamingo regulates epiboly and convergence/extension movements through cell cohesive and signalling functions during zebrafish gastrulation. Development. 2009;136:383–392. doi: 10.1242/dev.026542. PubMed DOI PMC

Katoh M. GIPC gene family (review) Int J Mol Med. 2002;9:585–589. PubMed

Sensoy O, Weinstein H. A mechanistic role of helix 8 in GPCRs: computational modeling of the dopamine D2 receptor interaction with the GIPC1-PDZ-domain. Biochim Biophys Acta. 1848;2015:976–983. PubMed PMC

Arango-Lievano M, Sensoy O, Borie A, Corbani M, Guillon G, Sokoloff P, Weinstein H, Jeanneteau F. A GIPC1-Palmitate switch modulates dopamine Drd3 receptor trafficking and signaling. Mol Cell Biol. 2016;36:1019–1031. doi: 10.1128/MCB.00916-15. PubMed DOI PMC

Jeanneteau F, Diaz J, Sokoloff P, Griffon N. Interactions of GIPC with dopamine D2, D3 but not D4 receptors define a novel mode of regulation of G protein-coupled receptors. Mol Biol Cell. 2004;15:696–705. doi: 10.1091/mbc.E03-05-0293. PubMed DOI PMC

Jeanneteau F, Guillin O, Diaz J, Griffon N, Sokoloff P. GIPC recruits GAIP (RGS19) to attenuate dopamine D2 receptor signaling. Mol Biol Cell. 2004;15:4926–4937. doi: 10.1091/mbc.E04-04-0285. PubMed DOI PMC

Kim J, Lee S, Ko S, Kim-Ha J. dGIPC is required for the locomotive activity and longevity in drosophila. Biochem Biophys Res Commun. 2010;402:565–570. doi: 10.1016/j.bbrc.2010.10.095. PubMed DOI

Choi I, Kim B, Byun JW, Baik SH, Huh YH, Kim JH, Mook-Jung I, Song WK, Shin JH, Seo H, et al. LRRK2 G2019S mutation attenuates microglial motility by inhibiting focal adhesion kinase. Nat Commun. 2015;6:8255. doi: 10.1038/ncomms9255. PubMed DOI PMC

Hannigan GE, Leung-Hagesteijn C, Fitz-Gibbon L, Coppolino MG, Radeva G, Filmus J, Bell JC, Dedhar S. Regulation of cell adhesion and anchorage-dependent growth by a new beta 1-integrin-linked protein kinase. Nature. 1996;379:91–96. doi: 10.1038/379091a0. PubMed DOI

Filipenko NR, Attwell S, Roskelley C, Dedhar S. Integrin-linked kinase activity regulates Rac- and Cdc42-mediated actin cytoskeleton reorganization via alpha-PIX. Oncogene. 2005;24:5837–5849. doi: 10.1038/sj.onc.1208737. PubMed DOI

Boulter E, Grall D, Cagnol S, Van Obberghen-Schilling E. Regulation of cell-matrix adhesion dynamics and Rac-1 by integrin linked kinase. FASEB J. 2006;20:1489–1491. doi: 10.1096/fj.05-4579fje. PubMed DOI

James NG, Digman MA, Gratton E, Barylko B, Ding X, Albanesi JP, Goldberg MS, Jameson DM. Number and brightness analysis of LRRK2 oligomerization in live cells. Biophys J. 2012;102:L41–L43. doi: 10.1016/j.bpj.2012.04.046. PubMed DOI PMC

Sakaguchi-Nakashima A, Meir JY, Jin Y, Matsumoto K, Hisamoto N. LRK-1, a C. elegans PARK8-related kinase, regulates axonal-dendritic polarity of SV proteins. Curr Biol. 2007;17:592–598. doi: 10.1016/j.cub.2007.01.074. PubMed DOI

Arranz AM, Delbroek L, Van Kolen K, Guimaraes MR, Mandemakers W, Daneels G, Matta S, Calafate S, Shaban H, Baatsen P, et al. LRRK2 functions in synaptic vesicle endocytosis through a kinase-dependent mechanism. J Cell Sci. 2015;128:541–552. doi: 10.1242/jcs.158196. PubMed DOI

Lee S, Imai Y, Gehrke S, Liu S, Lu B. The synaptic function of LRRK2. Biochem Soc Trans. 2012;40:1047–1051. doi: 10.1042/BST20120113. PubMed DOI

Parisiadou L, Yu J, Sgobio C, Xie C, Liu G, Sun L, Gu XL, Lin X, Crowley NA, Lovinger DM, Cai H. LRRK2 regulates synaptogenesis and dopamine receptor activation through modulation of PKA activity. Nat Neurosci. 2014;17:367–376. doi: 10.1038/nn.3636. PubMed DOI PMC

Piccoli G, Condliffe SB, Bauer M, Giesert F, Boldt K, De Astis S, Meixner A, Sarioglu H, Vogt-Weisenhorn DM, Wurst W, et al. LRRK2 controls synaptic vesicle storage and mobilization within the recycling pool. J Neurosci. 2011;31:2225–2237. doi: 10.1523/JNEUROSCI.3730-10.2011. PubMed DOI PMC

Borgs L, Peyre E, Alix P, Hanon K, Grobarczyk B, Godin JD, Purnelle A, Krusy N, Maquet P, Lefebvre P, et al. Dopaminergic neurons differentiating from LRRK2 G2019S induced pluripotent stem cells show early neuritic branching defects. Sci Rep. 2016;6:33377. doi: 10.1038/srep33377. PubMed DOI PMC

Shimizu K, Sato M, Tabata T. The Wnt5/planar cell polarity pathway regulates axonal development of the drosophila mushroom body neuron. J Neurosci. 2011;31:4944–4954. doi: 10.1523/JNEUROSCI.0154-11.2011. PubMed DOI PMC

Li X, Wang Y, Wang H, Liu T, Guo J, Yi W, Li Y. Epithelia-derived wingless regulates dendrite directional growth of drosophila ddaE neuron through the Fz-Fmi-Dsh-Rac1 pathway. Mol Brain. 2016;9:46. doi: 10.1186/s13041-016-0228-0. PubMed DOI PMC

Steimel A, Wong L, Najarro EH, Ackley BD, Garriga G, Hutter H. The flamingo ortholog FMI-1 controls pioneer-dependent navigation of follower axons in C. elegans. Development. 2010;137:3663–3673. doi: 10.1242/dev.054320. PubMed DOI PMC

Mrkusich EM, Flanagan DJ, Whitington PM. The core planar cell polarity gene prickle interacts with flamingo to promote sensory axon advance in the drosophila embryo. Dev Biol. 2011;358:224–230. doi: 10.1016/j.ydbio.2011.07.032. PubMed DOI

Tao H, Manak JR, Sowers L, Mei X, Kiyonari H, Abe T, Dahdaleh NS, Yang T, Wu S, Chen S, et al. Mutations in prickle orthologs cause seizures in flies, mice, and humans. Am J Hum Genet. 2011;88:138–149. doi: 10.1016/j.ajhg.2010.12.012. PubMed DOI PMC

Bassuk AG, Wallace RH, Buhr A, Buller AR, Afawi Z, Shimojo M, Miyata S, Chen S, Gonzalez-Alegre P, Griesbach HL, et al. A homozygous mutation in human PRICKLE1 causes an autosomal-recessive progressive myoclonus epilepsy-ataxia syndrome. Am J Hum Genet. 2008;83:572–581. doi: 10.1016/j.ajhg.2008.10.003. PubMed DOI PMC

Fox MH, Bassuk AG. PRICKLE1-related progressive Myoclonus epilepsy with ataxia. In: Pagon RA, Adam MP, Ardinger HH, Wallace SE, Amemiya A, LJH B, Bird TD, Fong CT, Mefford HC, RJH S, Stephens K, editors. GeneReviews(R) Seattle: University of Washington, Seattle University of Washington, Seattle; 1993.

Paemka L, Mahajan VB, Skeie JM, Sowers LP, Ehaideb SN, Gonzalez-Alegre P, Sasaoka T, Tao H, Miyagi A, Ueno N, et al. PRICKLE1 interaction with SYNAPSIN I reveals a role in autism spectrum disorders. PLoS One. 2013;8:e80737. doi: 10.1371/journal.pone.0080737. PubMed DOI PMC

Onishi K, Hollis E, Zou Y. Axon guidance and injury-lessons from Wnts and Wnt signaling. Curr Opin Neurobiol. 2014;27:232–240. doi: 10.1016/j.conb.2014.05.005. PubMed DOI PMC

Nagaoka T, Ohashi R, Inutsuka A, Sakai S, Fujisawa N, Yokoyama M, Huang YH, Igarashi M, Kishi M. The Wnt/planar cell polarity pathway component Vangl2 induces synapse formation through direct control of N-cadherin. Cell Rep. 2014;6:916–927. doi: 10.1016/j.celrep.2014.01.044. PubMed DOI

Chen J, Park CS, Tang SJ. Activity-dependent synaptic Wnt release regulates hippocampal long term potentiation. J Biol Chem. 2006;281:11910–11916. doi: 10.1074/jbc.M511920200. PubMed DOI

Farias GG, Alfaro IE, Cerpa W, Grabowski CP, Godoy JA, Bonansco C, Inestrosa NC. Wnt-5a/JNK signaling promotes the clustering of PSD-95 in hippocampal neurons. J Biol Chem. 2009;284:15857–15866. doi: 10.1074/jbc.M808986200. PubMed DOI PMC

Nixon-Abell J, Berwick DC, Granno S, Spain VA, Blackstone C, Harvey K. Protective LRRK2 R1398H variant enhances GTPase and Wnt signaling activity. Front Mol Neurosci. 2016;9:18. doi: 10.3389/fnmol.2016.00018. PubMed DOI PMC

Chan D, Citro A, Cordy JM, Shen GC, Wolozin B. Rac1 protein rescues neurite retraction caused by G2019S leucine-rich repeat kinase 2 (LRRK2) J Biol Chem. 2011;286:16140–16149. doi: 10.1074/jbc.M111.234005. PubMed DOI PMC

Lindqvist M, Horn Z, Bryja V, Schulte G, Papachristou P, Ajima R, Dyberg C, Arenas E, Yamaguchi TP, Lagercrantz H, Ringstedt T. Vang-like protein 2 and Rac1 interact to regulate adherens junctions. J Cell Sci. 2010;123:472–483. doi: 10.1242/jcs.048074. PubMed DOI PMC

Berwick DC, Javaheri B, Wetzel A, Hopkinson M, Nixon-Abell J, Granno S, Pitsillides AA, Harvey K. Pathogenic LRRK2 variants are gain-of-function mutations that enhance LRRK2-mediated repression of beta-catenin signaling. Mol Neurodegener. 2017;12:9. doi: 10.1186/s13024-017-0153-4. PubMed DOI PMC

Gao B, Song H, Bishop K, Elliot G, Garrett L, English MA, Andre P, Robinson J, Sood R, Minami Y, et al. Wnt signaling gradients establish planar cell polarity by inducing Vangl2 phosphorylation through Ror2. Dev Cell. 2011;20:163–176. doi: 10.1016/j.devcel.2011.01.001. PubMed DOI PMC

Kuss P, Kraft K, Stumm J, Ibrahim D, Vallecillo-Garcia P, Mundlos S, Stricker S. Regulation of cell polarity in the cartilage growth plate and perichondrium of metacarpal elements by HOXD13 and WNT5A. Dev Biol. 2014;385:83–93. doi: 10.1016/j.ydbio.2013.10.013. PubMed DOI

Lu C, Wan Y, Cao J, Zhu X, Yu J, Zhou R, Yao Y, Zhang L, Zhao H, Li H, et al. Wnt-mediated reciprocal regulation between cartilage and bone development during endochondral ossification. Bone. 2013;53:566–574. doi: 10.1016/j.bone.2012.12.016. PubMed DOI

Yang T, Bassuk AG, Fritzsch B. Prickle1 stunts limb growth through alteration of cell polarity and gene expression. Dev Dyn. 2013;242:1293–1306. doi: 10.1002/dvdy.24025. PubMed DOI PMC

Gandhi PN, Wang X, Zhu X, Chen SG, Wilson-Delfosse AL. The roc domain of leucine-rich repeat kinase 2 is sufficient for interaction with microtubules. J Neurosci Res. 2008;86:1711–1720. doi: 10.1002/jnr.21622. PubMed DOI PMC

Liu GH, Qu J, Suzuki K, Nivet E, Li M, Montserrat N, Yi F, Xu X, Ruiz S, Zhang W, et al. Progressive degeneration of human neural stem cells caused by pathogenic LRRK2. Nature. 2012;491:603–607. doi: 10.1038/nature11557. PubMed DOI PMC

Wang L, Xie C, Greggio E, Parisiadou L, Shim H, Sun L, Chandran J, Lin X, Lai C, Yang WJ, et al. The chaperone activity of heat shock protein 90 is critical for maintaining the stability of leucine-rich repeat kinase 2. J Neurosci. 2008;28:3384–3391. doi: 10.1523/JNEUROSCI.0185-08.2008. PubMed DOI PMC

Katoh M. Functional proteomics, human genetics and cancer biology of GIPC family members. Exp Mol Med. 2013;45:e26. doi: 10.1038/emm.2013.49. PubMed DOI PMC

Tan C, Deardorff MA, Saint-Jeannet JP, Yang J, Arzoumanian A, Klein PS. Kermit, a frizzled interacting protein, regulates frizzled 3 signaling in neural crest development. Development. 2001;128:3665–3674. PubMed

Ren N, Zhu C, Lee H, Adler PN. Gene expression during drosophila wing morphogenesis and differentiation. Genetics. 2005;171:625–638. doi: 10.1534/genetics.105.043687. PubMed DOI PMC

Djiane A, Mlodzik M. The drosophila GIPC homologue can modulate myosin based processes and planar cell polarity but is not essential for development. PLoS One. 2010;5:e11228. doi: 10.1371/journal.pone.0011228. PubMed DOI PMC

Grunewald TG, Pasedag SM, Butt E. Cell adhesion and transcriptional activity - defining the role of the novel Protooncogene LPP. Transl Oncol. 2009;2:107–116. doi: 10.1593/tlo.09112. PubMed DOI PMC

Ngan E, Northey JJ, Brown CM, Ursini-Siegel J, Siegel PM. A complex containing LPP and alpha-actinin mediates TGFbeta-induced migration and invasion of ErbB2-expressing breast cancer cells. J Cell Sci. 2013;126:1981–1991. doi: 10.1242/jcs.118315. PubMed DOI PMC

Petit MM, Meulemans SM, Alen P, Ayoubi TA, Jansen E, Van de Ven WJ. The tumor suppressor Scrib interacts with the zyxin-related protein LPP, which shuttles between cell adhesion sites and the nucleus. BMC Cell Biol. 2005;6:1. doi: 10.1186/1471-2121-6-1. PubMed DOI PMC

Vervenne HB, Crombez KR, Lambaerts K, Carvalho L, Koppen M, Heisenberg CP, Van de Ven WJ, Petit MM. Lpp is involved in Wnt/PCP signaling and acts together with Scrib to mediate convergence and extension movements during zebrafish gastrulation. Dev Biol. 2008;320:267–277. doi: 10.1016/j.ydbio.2008.05.529. PubMed DOI

Wu C, Keightley SY, Leung-Hagesteijn C, Radeva G, Coppolino M, Goicoechea S, McDonald JA, Dedhar S. Integrin-linked protein kinase regulates fibronectin matrix assembly, E-cadherin expression, and tumorigenicity. J Biol Chem. 1998;273:528–536. doi: 10.1074/jbc.273.1.528. PubMed DOI

Wu X, Wang J, Jiang H, Hu Q, Chen J, Zhang J, Zhu R, Liu W, Li B. Wnt3a activates beta1-integrin and regulates migration and adhesion of vascular smooth muscle cells. Mol Med Rep. 2014;9:1159–1164. PubMed

Lanni C, Necchi D, Pinto A, Buoso E, Buizza L, Memo M, Uberti D, Govoni S, Racchi M. Zyxin is a novel target for beta-amyloid peptide: characterization of its role in Alzheimer's pathogenesis. J Neurochem. 2013;125:790–799. doi: 10.1111/jnc.12154. PubMed DOI

Martynova NY, Ermolina LV, Ermakova GV, Eroshkin FM, Gyoeva FK, Baturina NS, Zaraisky AG. The cytoskeletal protein Zyxin inhibits Shh signaling during the CNS patterning in Xenopus laevis through interaction with the transcription factor Gli1. Dev Biol. 2013;380:37–48. doi: 10.1016/j.ydbio.2013.05.005. PubMed DOI

Luo S, Schaefer AM, Dour S, Nonet ML. The conserved LIM domain-containing focal adhesion protein ZYX-1 regulates synapse maintenance in Caenorhabditis elegans. Development. 2014;141:3922–3933. doi: 10.1242/dev.108217. PubMed DOI PMC

van Wijk NV, Witte F, Feike AC, Schambony A, Birchmeier W, Mundlos S, Stricker S. The LIM domain protein Wtip interacts with the receptor tyrosine kinase Ror2 and inhibits canonical Wnt signalling. Biochem Biophys Res Commun. 2009;390:211–216. doi: 10.1016/j.bbrc.2009.09.086. PubMed DOI

Hansen SD, Mullins RD. Lamellipodin promotes actin assembly by clustering Ena/VASP proteins and tethering them to actin filaments. Elife. 2015;4 PubMed PMC

Krause M, Leslie JD, Stewart M, Lafuente EM, Valderrama F, Jagannathan R, Strasser GA, Rubinson DA, Liu H, Way M, et al. Lamellipodin, an Ena/VASP ligand, is implicated in the regulation of lamellipodial dynamics. Dev Cell. 2004;7:571–583. doi: 10.1016/j.devcel.2004.07.024. PubMed DOI

Vehlow A, Soong D, Vizcay-Barrena G, Bodo C, Law AL, Perera U, Krause M. Endophilin, Lamellipodin, and Mena cooperate to regulate F-actin-dependent EGF-receptor endocytosis. EMBO J. 2013;32:2722–2734. doi: 10.1038/emboj.2013.212. PubMed DOI PMC

Tasaka G, Negishi M, Oinuma I. Semaphorin 4D/Plexin-B1-mediated M-Ras GAP activity regulates actin-based dendrite remodeling through Lamellipodin. J Neurosci. 2012;32:8293–8305. doi: 10.1523/JNEUROSCI.0799-12.2012. PubMed DOI PMC