Adhesion G Protein-coupled Receptor G1 (GPR56) Regulation by Sorting Nexin 27 Open Access

Iancu, Ariella Magen (2015)

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Adhesion G Protein-coupled Receptor G1 (G1), previously known as GPR56, plays a role in a wide variety of biological functions including cancer, immunology, and development. Mutations in G1 cause the brain developmental disorder Bilateral Fronto-Parietal Polymicrogyria (BFPP), and mutant forms of G1 associated with BFPP exhibit reduced cell surface expression and signaling. Sorting Nexin (SNX) family members typically modulate endosomal trafficking to the plasma membrane, and SNX27 has been recently identified as a potential modulator of G1 in a proteomic study. I showed that SNX27 interacts with both full-length G1 and N-terminally truncated G1 in HEK-293 cells. I also found that SNX27 increases total and surface expression of G1 and N-terminally truncated G1. Preliminary results indicate that SNX27 increases signaling of G1. Based on these results, SNX27 may be an important modulator of G1 expression and signaling in vivo.

Table of Contents

I. Introduction 1

A. G Protein-coupled Receptors 1

B. Adhesion GPCRs 3

C. Adhesion G Protein-coupled Receptor ADGR-G1 (G1) 5

D. Aims of This Thesis 12

II. Methods 13

A. Cell Culture 13

B. Plasmids 13

C. Antibodies 13

D. Western Blotting 14

E. Coimmunoprecipitation 14

F. Cell-Surface Biotinylation 14

G. Gene Reporter Assay: Dual-Glo Luciferase Assay 15

H. Statistical Analysis 16

III. Results 16

A. SNX27 Interacts with G1 16

B. Total and Surface Expression of G1 Increase with SNX27 17

C. SNX27 Increases G1 Signaling 20

IV. Discussion 21

V. Works Cited 24

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