title: Regulation of ROCKII membrane localization through its C-terminus.
abstract: RhoA activated kinases (ROCKs) are potent effectors of RhoA signaling for regulation of the cytoskeleton. ROCKs have been shown to be localized to several different subcellular locations, suggesting that its localization is context specific and regulated. However, the signaling mechanisms that control ROCK localization have not been clearly described. In this study we measured ROCKII localization following stimulation with the chemokine CXCL12 or adhesion to collagen 1. Strikingly, each of these extracellular signals targeted ROCKII to membrane protrusions. We further determined that both RhoA and PI3-kinase signaling are required for these stimuli to induce efficient membrane localization. Furthermore, we used a mutational approach to show that two separate domains predicted to respond to these localization signals, the Rho Binding Domain (RBD) and the Pleckstrin Homology domain (PH). Unexpectedly, we found that these two domains work synergistically to lead to membrane localization. This suggests a novel mechanism for controlling ROCKII localization at the membrane, in which the ROCKII C-terminus acts as a coincidence detector for spatial regulatory signals. In other words, efficient membrane targeting requires the ROCKII RBD to receive the RhoA signal and the PH domain to receive the phospholipid signal.
authors: Department of Pharmacology, LSU Health Sciences Center, New Orleans, LA 70119, USA.
authors: Swapnil S
authors: SS
authors: Kher
authors: Rebecca A
authors: RA
authors: Worthylake
mesh_terms: D022081:Cell Surface Extensions / Q000378:metabolism*; D054377:Chemokine CXCL12 / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D054460:rho-Associated Kinases / Q000378:metabolism*
chemical_list: C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D010743:Phospholipids; D003094:Collagen; D019869:Phosphatidylinositol 3-Kinases; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases