The tumor suppressor protein DLC1 maintains protein kinase D activity and Golgi secretory function

• Published in: The Journal of biological chemistry 2018-09, Vol.293 (37), p.14407-14416
• Main Authors: Jensch, Antje, Frey, Yannick, Bitschar, Katharina, Weber, Patrick, Schmid, Simone, Hausser, Angelika, Olayioye, Monilola A., Radde, Nicole E.
• Format: Article
• Language: English
• Subjects: Cell Biology; Cell Line, Tumor; Enzyme Activation; Exocytosis; GTPase-Activating Proteins - genetics; GTPase-Activating Proteins - metabolism; HEK293 Cells; Humans; Intracellular Membranes - metabolism; Models, Biological; Phosphorylation; Protein Kinase C - metabolism; rab GTP-Binding Proteins - metabolism; rho-Associated Kinases - metabolism; RNA Interference; Signal Transduction; Substrate Specificity; trans-Golgi Network - metabolism; Tumor Suppressor Proteins - genetics; Tumor Suppressor Proteins - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.RA118.003787

Many newly synthesized cellular proteins pass through the Golgi complex from where secretory transport carriers sort them to the plasma membrane and the extracellular environment. The formation of these secretory carriers at the trans-Golgi network is promoted by the protein kinase D (PKD) family of serine/threonine kinases. Here, using mathematical modeling and experimental validation of the PKD activation and substrate phosphorylation kinetics, we reveal that the expression level of the PKD substrate deleted in liver cancer 1 (DLC1), a Rho GTPase–activating protein that is inhibited by PKD-mediated phosphorylation, determines PKD activity at the Golgi membranes. RNAi-mediated depletion of DLC1 reduced PKD activity in a Rho–Rho-associated protein kinase (ROCK)–dependent manner, impaired the exocytosis of the cargo protein horseradish peroxidase, and was associated with the accumulation of the small GTPase RAB6 on Golgi membranes, indicating a protein-trafficking defect. In summary, our findings reveal that DLC1 maintains basal activation of PKD at the Golgi and Golgi secretory activity, in part by down-regulating Rho–ROCK signaling. We propose that PKD senses cytoskeletal changes downstream of DLC1 to coordinate Rho signaling with Golgi secretory function.