TY - JOUR
T1 - Deciphering protein kinase specificity through large-scale analysis of yeast phosphorylation site motifs
AU - Mok, Janine
AU - Kim, Philip M.
AU - Lam, Hugo Y.K.
AU - Piccirillo, Stacy
AU - Zhou, Xiuqiong
AU - Jeschke, Grace R.
AU - Sheridan, Douglas L.
AU - Parker, Sirlester A.
AU - Desai, Ved
AU - Jwa, Miri
AU - Cameroni, Elisabetta
AU - Niu, Hengyao
AU - Good, Matthew
AU - Remenyi, Attila
AU - Ma, Jia Lin Nianhan
AU - Sheu, Yi Jun
AU - Sassi, Holly E.
AU - Sopko, Richelle
AU - Chan, Clarence S.M.
AU - De Virgilio, Claudio
AU - Hollingsworth, Nancy M.
AU - Lim, Wendell A.
AU - Stern, David F.
AU - Stillman, Bruce
AU - Andrews, Brenda J.
AU - Gerstein, Mark B.
AU - Snyder, Michael
AU - Turk, Benjamin E.
PY - 2010/2/16
Y1 - 2010/2/16
N2 - Phosphorylation is a universal mechanism for regulating cell behavior in eukaryotes. Although protein kinases target short linear sequence motifs on their substrates, the rules for kinase substrate recognition are not completely understood. We used a rapid peptide screening approach to determine consensus phosphorylation site motifs targeted by 61 of the 122 kinases in Saccharomyces cerevisiae. By correlating these motifs with kinase primary sequence, we uncovered previously unappreciated rules for determining specificity within the kinase family, including a residue determining P-3 arginine specificity among members of the CMGC [CDK (cyclin-dependent kinase), MAPK (mitogen-activated protein kinase), GSK (glycogen synthase kinase), and CDK-like] group of kinases. Furthermore, computational scanning of the yeast proteome enabled the prediction of thousands of new kinasesubstrate relationships. We experimentally verified several candidate substrates of the Prk1 family of kinases in vitro and in vivo and identified a protein substrate of the kinase Vhs1. Together, these results elucidate how kinase catalytic domains recognize their phosphorylation targets and suggest general avenues for the identification of previously unknown kinase substrates across eukaryotes.
AB - Phosphorylation is a universal mechanism for regulating cell behavior in eukaryotes. Although protein kinases target short linear sequence motifs on their substrates, the rules for kinase substrate recognition are not completely understood. We used a rapid peptide screening approach to determine consensus phosphorylation site motifs targeted by 61 of the 122 kinases in Saccharomyces cerevisiae. By correlating these motifs with kinase primary sequence, we uncovered previously unappreciated rules for determining specificity within the kinase family, including a residue determining P-3 arginine specificity among members of the CMGC [CDK (cyclin-dependent kinase), MAPK (mitogen-activated protein kinase), GSK (glycogen synthase kinase), and CDK-like] group of kinases. Furthermore, computational scanning of the yeast proteome enabled the prediction of thousands of new kinasesubstrate relationships. We experimentally verified several candidate substrates of the Prk1 family of kinases in vitro and in vivo and identified a protein substrate of the kinase Vhs1. Together, these results elucidate how kinase catalytic domains recognize their phosphorylation targets and suggest general avenues for the identification of previously unknown kinase substrates across eukaryotes.
UR - https://www.scopus.com/pages/publications/77952986553
U2 - 10.1126/scisignal.2000482
DO - 10.1126/scisignal.2000482
M3 - Article
C2 - 20159853
AN - SCOPUS:77952986553
SN - 1945-0877
VL - 3
SP - ra12
JO - Science Signaling
JF - Science Signaling
IS - 109
ER -