Engineering allosteric regulation in protein kinases

David Pincus, Jai P. Pandey, Zoë A. Feder, Pau Creixell, Orna Resnekov, Kimberly A Reynolds

Research output: Contribution to journalArticle

Abstract

Phosphoregulation, in which the addition of a negatively charged phosphate group modulates protein activity, enables dynamic cellular responses. To understand how new phosphoregulation might be acquired, we mutationally scanned the surface of a prototypical yeast kinase (Kss1) to identify potential regulatory sites. The data revealed a set of spatially distributed “hotspots” that might have coevolved with the active site and preferentially modulated kinase activity. By engineering simple consensus phosphorylation sites at these hotspots, we rewired cell signaling in yeast. Using the same approach with a homolog yeast mitogen-activated protein kinase, Hog1, we introduced new phosphoregulation that modified its localization and signaling dynamics. Beyond revealing potential use in synthetic biology, our findings suggest that the identified hotspots contribute to the diversity of natural allosteric regulatory mechanisms in the eukaryotic kinome and, given that some are mutated in cancers, understanding these hotspots may have clinical relevance to human disease.

Original languageEnglish (US)
Article numbereaar3250
JournalScience Signaling
Volume11
Issue number555
DOIs
StatePublished - Nov 6 2018

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Allosteric Regulation
Yeast
Protein Kinases
Yeasts
Phosphotransferases
Synthetic Biology
Cell signaling
Phosphorylation
Mitogen-Activated Protein Kinases
Catalytic Domain
Phosphates
Neoplasms
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Pincus, D., Pandey, J. P., Feder, Z. A., Creixell, P., Resnekov, O., & Reynolds, K. A. (2018). Engineering allosteric regulation in protein kinases. Science Signaling, 11(555), [eaar3250]. https://doi.org/10.1126/scisignal.aar3250

Engineering allosteric regulation in protein kinases. / Pincus, David; Pandey, Jai P.; Feder, Zoë A.; Creixell, Pau; Resnekov, Orna; Reynolds, Kimberly A.

In: Science Signaling, Vol. 11, No. 555, eaar3250, 06.11.2018.

Research output: Contribution to journalArticle

Pincus, D, Pandey, JP, Feder, ZA, Creixell, P, Resnekov, O & Reynolds, KA 2018, 'Engineering allosteric regulation in protein kinases', Science Signaling, vol. 11, no. 555, eaar3250. https://doi.org/10.1126/scisignal.aar3250
Pincus D, Pandey JP, Feder ZA, Creixell P, Resnekov O, Reynolds KA. Engineering allosteric regulation in protein kinases. Science Signaling. 2018 Nov 6;11(555). eaar3250. https://doi.org/10.1126/scisignal.aar3250
Pincus, David ; Pandey, Jai P. ; Feder, Zoë A. ; Creixell, Pau ; Resnekov, Orna ; Reynolds, Kimberly A. / Engineering allosteric regulation in protein kinases. In: Science Signaling. 2018 ; Vol. 11, No. 555.
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