2. Academic Validation
  3. Systematic proteomic analysis of LRRK2-mediated Rab GTPase phosphorylation establishes a connection to ciliogenesis

Systematic proteomic analysis of LRRK2-mediated Rab GTPase phosphorylation establishes a connection to ciliogenesis

  • Elife. 2017 Nov 10;6:e31012. doi: 10.7554/eLife.31012.
Martin Steger # 1,  Federico Diez # 2,  Herschel S Dhekne 3,  Pawel Lis 2,  Raja S Nirujogi 2,  Ozge Karayel 1,  Francesca Tonelli 2,  Terina N Martinez 4,  Esben Lorentzen 5,  Suzanne R Pfeffer 3,  Dario R Alessi 2,  Matthias Mann 1
Affiliations

Affiliations

  • 1 Department of Proteomics and Signal Transduction, Max-Planck-Institute of Biochemistry, Martinsried, Germany.
  • 2 Medical Research Council Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee, United Kingdom.
  • 3 Department of Biochemistry, Stanford University School of Medicine, Stanford, United States.
  • 4 The Michael J. Fox Foundation for Parkinson's Research, New York, United States.
  • 5 Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
  • # Contributed equally.
PMID: 29125462 DOI: 10.7554/eLife.31012
Abstract

We previously reported that Parkinson's disease (PD) kinase LRRK2 phosphorylates a subset of Rab GTPases on a conserved residue in their switch-II domains (Steger et al., 2016) (PMID: 26824392). Here, we systematically analyzed the Rab protein family and found 14 of them (Rab3A/B/C/D, Rab5A/B/C, Rab8A/B, Rab10, Rab12, Rab29, Rab35 and Rab43) to be specifically phosphorylated by LRRK2, with evidence for endogenous phosphorylation for ten of them (Rab3A/B/C/D, Rab8A/B, Rab10, Rab12, Rab35 and Rab43). Affinity enrichment mass spectrometry revealed that the primary ciliogenesis regulator, RILPL1 specifically interacts with the LRRK2-phosphorylated forms of Rab8A and Rab10, whereas RILPL2 binds to phosphorylated Rab8A, Rab10, and Rab12. Induction of primary cilia formation by serum starvation led to a two-fold reduction in ciliogenesis in fibroblasts derived from pathogenic LRRK2-R1441G knock-in mice. These results implicate LRRK2 in primary ciliogenesis and suggest that Rab-mediated protein transport and/or signaling defects at cilia may contribute to LRRK2-dependent pathologies.

Keywords

LRRK2; Parkinson's disease; Rab GTPases; biochemistry; ciliogenesis; human; mass spectrometry; mouse; proteomics.

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