Comprehensive profiling of the STE20 kinase family defines features essential for selective substrate targeting and signaling output.

Autor: Miller CJ; Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, United States of America., Lou HJ; Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, United States of America., Simpson C; Biotech Research and Innovation Center, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark., van de Kooij B; Departments of Biological Engineering and Biology, MIT Center for Precision Cancer Medicine and Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America., Ha BH; Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, United States of America., Fisher OS; Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, United States of America., Pirman NL; Department of Cellular and Molecular Physiology and Systems Biology Institute, Yale University, New Haven, Connecticut, United States of America., Boggon TJ; Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, United States of America., Rinehart J; Department of Cellular and Molecular Physiology and Systems Biology Institute, Yale University, New Haven, Connecticut, United States of America., Yaffe MB; Departments of Biological Engineering and Biology, MIT Center for Precision Cancer Medicine and Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America., Linding R; Biotech Research and Innovation Center, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark., Turk BE; Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, United States of America.
Jazyk: angličtina
Zdroj: PLoS biology [PLoS Biol] 2019 Mar 21; Vol. 17 (3), pp. e2006540. Date of Electronic Publication: 2019 Mar 21 (Print Publication: 2019).
DOI: 10.1371/journal.pbio.2006540
Abstrakt: Specificity within protein kinase signaling cascades is determined by direct and indirect interactions between kinases and their substrates. While the impact of localization and recruitment on kinase-substrate targeting can be readily assessed, evaluating the relative importance of direct phosphorylation site interactions remains challenging. In this study, we examine the STE20 family of protein serine-threonine kinases to investigate basic mechanisms of substrate targeting. We used peptide arrays to define the phosphorylation site specificity for the majority of STE20 kinases and categorized them into four distinct groups. Using structure-guided mutagenesis, we identified key specificity-determining residues within the kinase catalytic cleft, including an unappreciated role for the kinase β3-αC loop region in controlling specificity. Exchanging key residues between the STE20 kinases p21-activated kinase 4 (PAK4) and Mammalian sterile 20 kinase 4 (MST4) largely interconverted their phosphorylation site preferences. In cells, a reprogrammed PAK4 mutant, engineered to recognize MST substrates, failed to phosphorylate PAK4 substrates or to mediate remodeling of the actin cytoskeleton. In contrast, this mutant could rescue signaling through the Hippo pathway in cells lacking multiple MST kinases. These observations formally demonstrate the importance of catalytic site specificity for directing protein kinase signal transduction pathways. Our findings further suggest that phosphorylation site specificity is both necessary and sufficient to mediate distinct signaling outputs of STE20 kinases and imply broad applicability to other kinase signaling systems.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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