Research Project
9516572
RAF kinases, central to the MAPK pathway, are highly implicated in metastatic melanomas and represent an established drug target. However, the current pharmacological intervention with RAF inhibitors exhibits adverse effects including intrinsic drug resistance and the development of secondary malignancies. RAF kinases have kinase domain and regulatory domains that are jointly involved in regulation, substrate recognition and catalysis. We will apply biochemistry, structural biology, and biophysics to delineate the regulatory mechanisms of the full-length RAF kinases in different genetic backgrounds. The Specific Aims of the proposal are to (1) Discover how regulatory domains coordinate to activate wild type BRAF. (2) Elucidate how the ?loss- of-function? BRAF mutants (impaired or ?kinase-dead?) gain enhanced ability to drive tumors in concert with CRAF. We will characterize the full-length BRAFkinase-dead/CRAF to understand how ?kinase-dead? mutations affect kinase function. (3) Illuminate how ?gain-of-function? P-loop mutation aberrantly activates BRAF. Our goal is to understand the regulation mechanisms of RAF enzymes to facilitate discovery of effective and specific inhibitors to block the disease-associated RAF kinases.
