Functional analysis of protein kinase networks in living cells: beyond "knock-outs" and "knock-downs".
The identification of over 500 protein kinases encoded by the human genome sequence offers one measure of the importance of protein kinase networks in cell biology. High throughput technologies for inactivating genes are producing an awe-inspiring amount of data on the cellular and organismal effects of reducing the levels of individual protein kinases. Despite these technical advances, our understanding of kinase networks remains imprecise. Major challenges include correctly assigning kinases to particular networks, understanding how they are regulated, and identifying the relevant in vivo substrates. Genetic methods provide a way of addressing these questions, but their application requires understanding the nuances of how different types of mutations can affect protein kinases. The goal of this article is to provide a brief introductory primer into these issues using examples from yeast MAPK cascades and to motivate future systematic genetic analysis focusing on individual residues of protein kinases.
Madhani
Hiten D
HD
Department of Biochemistry and Biophysics, University of California, 600 16th St., San Fransico, CA 94143-2200, USA. hiten@biochem.usf.edu
D005821:Genetic Techniques; D047908:Intracellular Signaling Peptides and Proteins; D020930:MAP Kinase Kinase Kinases / Q000235:genetics / Q000378:metabolism; D020935:MAP Kinase Signaling System; D020929:Mitogen-Activated Protein Kinase Kinases; D020928:Mitogen-Activated Protein Kinases / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015003:Yeasts / Q000235:genetics
D047908:Intracellular Signaling Peptides and Proteins; D029701:Saccharomyces cerevisiae Proteins; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; C075580:FUS3 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases; C078754:STE20 protein, S cerevisiae; C067018:Ste11 protein, S cerevisiae; D020929:Mitogen-Activated Protein Kinase Kinases; C119844:STE7 protein, S cerevisiae