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Differential protein–protein interactions of LRRK 1 and LRRK 2 indicate roles in distinct cellular signaling pathways
Author(s) -
Reyniers Lauran,
Del Giudice Maria Grazia,
Civiero Laura,
Belluzzi Elisa,
Lobbestael Evy,
Beilina Alexandra,
Arrigoni Giorgio,
Derua Rita,
Waelkens Etienne,
Li Yan,
Crosio Claudia,
Iaccarino Ciro,
Cookson Mark R.,
Baekelandt Veerle,
Greggio Elisa,
Taymans JeanMarc
Publication year - 2014
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/jnc.12798
Subject(s) - biology , microbiology and biotechnology , signal transduction
Genetic studies show that LRRK 2, and not its closest paralogue LRRK 1, is linked to Parkinson's disease. To gain insight into the molecular and cellular basis of this discrepancy, we searched for LRRK 1‐ and LRRK 2‐specific cellular processes by identifying their distinct interacting proteins. A protein microarray‐based interaction screen was performed with recombinant 3xFlag‐ LRRK 1 and 3xFlag‐ LRRK 2 and, in parallel, co‐immunoprecipitation followed by mass spectrometry was performed from SH ‐ SY 5Y neuroblastoma cell lines stably expressing 3xFlag‐ LRRK 1 or 3xFlag‐ LRRK 2. We identified a set of LRRK 1‐ and LRRK 2‐specific as well as common interactors. One of our most prominent findings was that both screens pointed to epidermal growth factor receptor ( EGF ‐R) as a LRRK 1‐specific interactor, while 14‐3‐3 proteins were LRRK 2‐specific. This is consistent with phosphosite mapping of LRRK 1, revealing phosphosites outside of 14‐3‐3 consensus binding motifs. To assess the functional relevance of these interactions, SH ‐ SY 5Y‐ LRRK 1 and ‐ LRRK 2 cell lines were treated with LRRK 2 kinase inhibitors that disrupt 14‐3‐3 binding, or with EGF , an EGF ‐R agonist. Redistribution of LRRK 2, not LRRK 1, from diffuse cytoplasmic to filamentous aggregates was observed after inhibitor treatment. Similarly, EGF induced translocation of LRRK 1, but not of LRRK 2, to endosomes. Our study confirms that LRRK 1 and LRRK 2 can carry out distinct functions by interacting with different cellular proteins.LRRK1 and LRRK2 (leucine‐rich repeat kinase) interaction partners were identified by two different protein‐protein interaction screens. These confirmed epidermal growth factor receptor (EGR‐R) as a LRRK1‐specific interactor, while 14‐3‐3 proteins were LRRK2‐specific. Functional analysis of these interactions and the pathways they mediate shows that LRRK1 and LRRK2 signaling do not intersect, reflective of the differential role of both LRRKs in Parkinson's disease.