The proposed project seeks to characterize a novel class of calcium/calmodulin-dependent protein kinases. This protein family is the CAMGUKs, a subgroup of the MAGUKs, the membrane-associated guanylate kinases. The MAGUKs have been shown to organize cell-cell contacts, and be responsible for the control of cellular proliferation. The CAMGUK members are MAGUK proteins with an amino-terminal extension that has substantial homology to the multifunctional calcium/calmodulin-dependent protein kinase (CaMK II). The function of CAMGUK is unknown, though it has been shown to be essential for vulval induction in C. elegans. For this project, the tissue distribution and expression, plus the intracellular localization will be examined using Western and Northern blotting methods and immunocytochemistry. Further, CAMGUK will be purified and its calmodulin-binding and enzymatic properties determined, including those of the CaMKII domain. Site-directed mutagenesis of CAMGUK, followed by expression, purification, and characterization of mutant proteins, will be carried out in order to elucidate the functionality of the domains of the CaMK lI homologous region.
