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Structural Basis for Rab8a GTPase Recruitment of RILPL2 via LRRK2-Mediated Phosphorylation of Switch 2

41 Pages Posted: 11 Sep 2019 Sneak Peek Status: Review Complete

See all articles by Dieter Waschbusch

Dieter Waschbusch

Trinity College (Dublin) - School of Biochemistry and Immunology

Elena Purlyte

University of Dundee - MRC Protein Phosphorylation and Ubiquitylation Unit

Prosenjit Pal

University of Dundee - MRC Protein Phosphorylation and Ubiquitylation Unit

Emma McGrath

Trinity College (Dublin) - School of Biochemistry and Immunology

Dario R. Alessi

University of Dundee - MRC Protein Phosphorylation and Ubiquitylation Unit

Amir Rafiq Khan

Trinity College (Dublin) - School of Biochemistry and Immunology

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Abstract

Rab8a GTPase is associated with the dynamic regulation of membrane protrusions in polarized cells.  Rab8a is one of several Rab-family GTPases that are substrates of leucine-rich repeat kinase 2 (LRRK2), a serine/threonine kinase that is linked to inherited Parkinson's disease.  Rab8a is phosphorylated at T72 (pT72) in its switch 2 helix and the post-translational modification facilitates phospho-Rab8a (pRab8a) interactions with RILPL2, which subsequently regulates ciliogenesis. Here we report the crystal structure of pRab8a in complex with the phospho-Rab binding domain of RILPL2.  The complex is a heterotetramer with RILPL2 forming a central alpha-helical dimer that bridges two pRab8a molecules.  The N-termini of the a-helices cross over to form an X-shaped cap (X-cap) that enables electrostatic interactions between Arg residues from RILPL2 and the phosphate moiety from pT72.  RILPL2 residues in the X-cap that are critical for pRab8a binding are conserved in the RILP family of effector proteins.  These structural insights revealed that JIP3 and JIP4 also possess a phospho-Rab binding domain similar to RILPL2 including the conserved residues that bind to pRab8A. This prompted us to analyze these proteins and we found that JIP3 and JIP4 interact specifically with LRRK2-phosphorylated Rab10, suggesting a general mode of recognition for phosphorylated Rab GTPases by phospho-specific effectors.

Keywords: Rab8a GTPase, LRRK2 protein kinase, phosphorylation, membrane trafficking, Parkinson's disease, RILP-like protein 2, X-ray crystallography

Suggested Citation

Waschbusch, Dieter and Purlyte, Elena and Pal, Prosenjit and McGrath, Emma and Alessi, Dario R. and Khan, Amir Rafiq, Structural Basis for Rab8a GTPase Recruitment of RILPL2 via LRRK2-Mediated Phosphorylation of Switch 2 (September 10, 2019). STRUCTURE-D-19-00262. Available at SSRN: https://ssrn.com/abstract=3451451 or http://dx.doi.org/10.2139/ssrn.3451451
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Dieter Waschbusch

Trinity College (Dublin) - School of Biochemistry and Immunology

2-3 College Green
Dublin, Leinster
Ireland

Elena Purlyte

University of Dundee - MRC Protein Phosphorylation and Ubiquitylation Unit

United Kingdom

Prosenjit Pal

University of Dundee - MRC Protein Phosphorylation and Ubiquitylation Unit

United Kingdom

Emma McGrath

Trinity College (Dublin) - School of Biochemistry and Immunology

2-3 College Green
Dublin, Leinster
Ireland

Dario R. Alessi

University of Dundee - MRC Protein Phosphorylation and Ubiquitylation Unit

United Kingdom

Amir Rafiq Khan (Contact Author)

Trinity College (Dublin) - School of Biochemistry and Immunology ( email )

2-3 College Green
Dublin, Leinster
Ireland

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