• Register
X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

X

Leaving Community

Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.

No
Yes

14-3-3 binding to LRRK2 is disrupted by multiple Parkinson's disease-associated mutations and regulates cytoplasmic localization.

LRRK2 (leucine-rich repeat protein kinase 2) is mutated in a significant number of Parkinson's disease patients, but still little is understood about how it is regulated or functions. In the present study we have demonstrated that 14-3-3 protein isoforms interact with LRRK2. Consistent with this, endogenous LRRK2 isolated from Swiss 3T3 cells or various mouse tissues is associated with endogenous 14-3-3 isoforms. We have established that 14-3-3 binding is mediated by phosphorylation of LRRK2 at two conserved residues (Ser910 and Ser935) located before the leucine-rich repeat domain. Our results suggests that mutation of Ser910 and/or Ser935 to disrupt 14-3-3 binding does not affect intrinsic protein kinase activity, but induces LRRK2 to accumulate within discrete cytoplasmic pools, perhaps resembling inclusion bodies. To investigate links between 14-3-3 binding and Parkinson's disease, we studied how 41 reported mutations of LRRK2 affected 14-3-3 binding and cellular localization. Strikingly, we found that five of the six most common pathogenic mutations (R1441C, R1441G, R1441H, Y1699C and I2020T) display markedly reduced phosphorylation of Ser910/Ser935 thereby disrupting interaction with 14-3-3. We have also demonstrated that Ser910/Ser935 phosphorylation and 14-3-3 binding to endogenous LRRK2 is significantly reduced in tissues of homozygous LRRK2(R1441C) knock-in mice. Consistent with 14-3-3 regulating localization, all of the common pathogenic mutations displaying reduced 14-3-3-binding accumulated within inclusion bodies. We also found that three of the 41 LRRK2 mutations analysed displayed elevated protein kinase activity (R1728H, ~2-fold; G2019S, ~3-fold; and T2031S, ~4-fold). These results provide the first evidence suggesting that 14-3-3 regulates LRRK2 and that disruption of the interaction of LRRK2 with 14-3-3 may be linked to Parkinson's disease.

Pubmed ID: 20642453

Authors

  • Nichols RJ
  • Dzamko N
  • Morrice NA
  • Campbell DG
  • Deak M
  • Ordureau A
  • Macartney T
  • Tong Y
  • Shen J
  • Prescott AR
  • Alessi DR

Journal

The Biochemical journal

Publication Data

September 15, 2010

Associated Grants

  • Agency: Medical Research Council, Id: G0700656
  • Agency: Medical Research Council, Id: MC_U127070193
  • Agency: Medical Research Council, Id:

Mesh Terms

  • 14-3-3 Proteins
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Brain
  • Cell Line
  • Cytoplasm
  • Green Fluorescent Proteins
  • Humans
  • Immunoprecipitation
  • Kidney
  • Mice
  • Mice, Knockout
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Mutation
  • Parkinson Disease
  • Phosphorylation
  • Protein Binding
  • Protein-Serine-Threonine Kinases
  • Sequence Homology, Amino Acid
  • Serine
  • Spleen
  • Swiss 3T3 Cells