title: Exocyst function is regulated by effector phosphorylation.
abstract: The exocyst complex tethers vesicles at sites of fusion through interactions with small GTPases. The G protein RalA resides on Glut4 vesicles, and binds to the exocyst after activation by insulin, but must then disengage to ensure continuous exocytosis. Here we report that, after recognition of the exocyst by activated RalA, disengagement occurs through phosphorylation of its effector Sec5, rather than RalA inactivation. Sec5 undergoes phosphorylation in the G-protein binding domain, allosterically reducing RalA interaction. The phosphorylation event is catalysed by protein kinase C and is reversed by an exocyst-associated phosphatase. Introduction of Sec5 bearing mutations of the phosphorylation site to either alanine or aspartate disrupts insulin-stimulated Glut4 exocytosis, as well as other trafficking processes in polarized epithelial cells and during development of zebrafish embryos. The exocyst thus serves as a 'gatekeeper' for exocytic vesicles through a circuit of engagement, disengagement and re-engagement with G proteins.
authors.affiliation: Life Sciences Institute, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA.
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Chen
authors.forename: Dara
authors.initials: D
authors.lastname: Leto
authors.forename: Junyu
authors.initials: J
authors.lastname: Xiao
authors.forename: John
authors.initials: J
authors.lastname: Goss
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.forename: Jordan A
authors.initials: JA
authors.lastname: Shavit
authors.forename: Tingting
authors.initials: T
authors.lastname: Xiong
authors.forename: Genggeng
authors.initials: G
authors.lastname: Yu
authors.forename: David
authors.initials: D
authors.lastname: Ginsburg
authors.forename: Derek
authors.initials: D
authors.lastname: Toomre
authors.forename: Zhaohui
authors.initials: Z
authors.lastname: Xu
authors.forename: Alan R
authors.initials: AR
authors.lastname: Saltiel
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D055162:Biocatalysis; D005089:Exocytosis; D020558:GTP Phosphohydrolases / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D015027:Zebrafish
chemical_list: D011493:Protein Kinase C; D020558:GTP Phosphohydrolases