Centrosomal anchoring of protein kinase C betaII by pericentrin controls microtubule organization, spindle function, and cytokinesis.
Location is a critical determinant in dictating the cellular function of protein kinase C (PKC). Scaffold proteins contribute to localization by poising PKC at specific intracellular sites. Using a yeast two-hybrid screen, we identified the centrosomal protein pericentrin as a scaffold that tethers PKC betaII to centrosomes. Co-immunoprecipitation studies reveal that the native proteins interact in cells. Co-overexpression studies show that the interaction is mediated by the C1A domain of PKC and a segment of pericentrin within residues 494-593. Immunofluorescence analysis reveals that endogenous PKC betaII colocalizes with pericentrin at centrosomes. Disruption of this interaction by expression of the interacting region of pericentrin results in release of PKC from the centrosome, microtubule disorganization, and cytokinesis failure. Overexpression of this disrupting fragment has no effect in cells lacking PKC betaII, indicating a specific regulatory role of this isozyme in centrosome function. These results reveal a novel role for PKC betaII in cytokinesis and indicate that this function is mediated by an interaction with pericentrin at centrosomes.
Chen
Dan
D
Department of Pharmacology, School of Medicine, University of California at San Diego, La Jolla, California 92093-0640, USA.
Purohit
Aruna
A
Halilovic
Ensar
E
Doxsey
Stephen J
SJ
Newton
Alexandra C
AC
D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002455:Cell Division; D002460:Cell Line; D018385:Centrosome / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta; D051381:Rats; D008941:Spindle Apparatus / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D000941:Antigens; D002352:Carrier Proteins; C086178:pericentrin; D011493:Protein Kinase C; D064546:Protein Kinase C beta