GPR107, a G-protein-coupled Receptor Essential for Intoxication by Pseudomonas aeruginosa Exotoxin A, Localizes to the Golgi and Is Cleaved by Furin

• Published in: The Journal of biological chemistry 2014-08, Vol.289 (35), p.24005-24018
• Main Authors: Tafesse, Fikadu G., Guimaraes, Carla P., Maruyama, Takeshi, Carette, Jan E., Lory, Stephen, Brummelkamp, Thijn R., Ploegh, Hidde L.
• Format: Article
• Language: English
• Subjects: ADP Ribose Transferases - genetics; ADP Ribose Transferases - toxicity; Bacterial Toxins - genetics; Bacterial Toxins - toxicity; Base Sequence; Cell Biology; DNA Primers; Endocytosis; Exotoxins - genetics; Exotoxins - toxicity; Furin - metabolism; Mutation; Polymerase Chain Reaction; Protein Transport; Proteolysis; Pseudomonas aeruginosa Exotoxin A; Receptors, G-Protein-Coupled - metabolism; Receptors, G-Protein-Coupled - physiology; trans-Golgi Network - metabolism; Virulence Factors - genetics; Virulence Factors - toxicity
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M114.589275

A number of toxins, including exotoxin A (PE) of Pseudomonas aeruginosa, kill cells by inhibiting protein synthesis. PE kills by ADP-ribosylation of the translation elongation factor 2, but many of the host factors required for entry, membrane translocation, and intracellular transport remain to be elucidated. A genome-wide genetic screen in human KBM7 cells was performed to uncover host factors used by PE, several of which were confirmed by CRISPR/Cas9-gene editing in a different cell type. Several proteins not previously implicated in the PE intoxication pathway were identified, including GPR107, an orphan G-protein-coupled receptor. GPR107 localizes to the trans-Golgi network and is essential for retrograde transport. It is cleaved by the endoprotease furin, and a disulfide bond connects the two cleaved fragments. Compromising this association affects the function of GPR107. The N-terminal region of GPR107 is critical for its biological function. GPR107 might be one of the long-sought receptors that associates with G-proteins to regulate intracellular vesicular transport.