RT Journal Article SR Electronic T1 Structural basis for Rab8a GTPase recruitment of RILPL2 via LRRK2 phosphorylation of switch 2 JF bioRxiv FD Cold Spring Harbor Laboratory SP 739813 DO 10.1101/739813 A1 Waschbüsch, Dieter A1 Purlyte, Elena A1 Pal, Prosenjit A1 McGrath, Emma A1 Alessi, Dario R. A1 Khan, Amir R. YR 2019 UL http://biorxiv.org/content/early/2019/08/19/739813.abstract AB 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 α-helical dimer that bridges two pRab8a molecules. The N-termini of the α-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. We find that JIP3 and JIP4 also interact specifically with LRRK2-phosphorylated Rab10, suggesting a general mode of recognition for phosphorylated Rab GTPases by phospho-specific effectors.