Systematic Discovery of Short Linear Motifs Decodes Calcineurin Phosphatase Signaling.
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Date
2020-07-16ICR Author
Author
Wigington, CP
Roy, J
Damle, NP
Yadav, VK
Blikstad, C
Resch, E
Wong, CJ
Mackay, DR
Wang, JT
Krystkowiak, I
Bradburn, DA
Tsekitsidou, E
Hong, SH
Kaderali, MA
Xu, S-L
Stearns, T
Gingras, A-C
Ullman, KS
Ivarsson, Y
Davey, NE
Cyert, MS
Type
Journal Article
Metadata
Show full item recordAbstract
Short linear motifs (SLiMs) drive dynamic protein-protein interactions essential for signaling, but sequence degeneracy and low binding affinities make them difficult to identify. We harnessed unbiased systematic approaches for SLiM discovery to elucidate the regulatory network of calcineurin (CN)/PP2B, the Ca2+-activated phosphatase that recognizes LxVP and PxIxIT motifs. In vitro proteome-wide detection of CN-binding peptides, in vivo SLiM-dependent proximity labeling, and in silico modeling of motif determinants uncovered unanticipated CN interactors, including NOTCH1, which we establish as a CN substrate. Unexpectedly, CN shows SLiM-dependent proximity to centrosomal and nuclear pore complex (NPC) proteins-structures where Ca2+ signaling is largely uncharacterized. CN dephosphorylates human and yeast NPC proteins and promotes accumulation of a nuclear transport reporter, suggesting conserved NPC regulation by CN. The CN network assembled here provides a resource to investigate Ca2+ and CN signaling and demonstrates synergy between experimental and computational methods, establishing a blueprint for examining SLiM-based networks.
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Subject
Hela Cells
Centrosome
Humans
Saccharomyces cerevisiae
Phosphoric Monoester Hydrolases
Calcineurin
Nuclear Pore Complex Proteins
Saccharomyces cerevisiae Proteins
Proteome
Biotinylation
Signal Transduction
Amino Acid Motifs
Active Transport, Cell Nucleus
Phosphorylation
Computer Simulation
Receptor, Notch1
Mass Spectrometry
HEK293 Cells
Protein Interaction Maps
Language
eng
Date accepted
2020-05-26
License start date
2020-07-08
Citation
Molecular cell, 2020, 79 (2), pp. 342 - 358.e12
Publisher
CELL PRESS