Title: | Identification of p21-activated kinase specificity determinants in budding yeast: a single amino acid substitution imparts Ste20 specificity to Cla4 |
Address: | "Department of Biology and Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229, USA" |
DOI: | 10.1128/MCB.23.5.1569-1580.2003 |
ISSN/ISBN: | 0270-7306 (Print) 1098-5549 (Electronic) 0270-7306 (Linking) |
Abstract: | "Two closely related p21-activated kinases from Saccharomyces cerevisiae, Ste20 and Cla4, interact with and are regulated by Cdc42, a small Rho-like GTPase. These kinases are argued to perform a common essential function, based on the observation that the single mutants are viable whereas the double mutant is inviable. Despite having a common upstream regulator and at least one common function, these molecules also have many distinct cellular signaling roles. Ste20 signals upstream of several mitogen-activated protein kinase cascades (e.g., pheromone response, filamentous growth, and high osmolarity), and Cla4 signals during budding and cytokinesis. In order to investigate how these kinases are directed to distinct functions, we sought to identify specificity determinants within Ste20 and Cla4. To this end, we constructed both chimeric fusions and point mutants and tested their ability to perform unique and shared cellular roles. Specificity determinants for both kinases were mapped to the C-terminal kinase domains. Remarkably, the substitution of a single amino acid, threonine 818, from Ste20 into an otherwise wild-type Cla4, Cla4D772T, conferred the ability to perform many Ste20-specific functions" |
Keywords: | "Blotting, Western Glutathione Transferase/metabolism Haploidy Intracellular Signaling Peptides and Proteins MAP Kinase Kinase Kinases/metabolism Phosphorylation Plasmids/metabolism Point Mutation Protein Binding Protein Serine-Threonine Kinases/*metabolis;" |
Notes: | "MedlineKeniry, Megan E Sprague, George F Jr eng R01 GM030027/GM/NIGMS NIH HHS/ T32 GM007413/GM/NIGMS NIH HHS/ GM-30027/GM/NIGMS NIH HHS/ GM07413/GM/NIGMS NIH HHS/ Research Support, U.S. Gov't, P.H.S. 2003/02/18 Mol Cell Biol. 2003 Mar; 23(5):1569-80. doi: 10.1128/MCB.23.5.1569-1580.2003" |