Glucocorticoid receptor Thr524 phosphorylation by MINK1 induces interactions with 14-3-3 protein regulators

DOI: 10.1016/j.jbc.2021.100551

Authors: Claire C. Munier (AstraZeneca; Technische Universiteit Eindhoven) , Leonardo De Maria (AstraZeneca) , Karl Edman (AstraZeneca) , Anders Gunnarsson (AstraZeneca) , Marianna Longo (University of Dundee) , Carol Mackintosh (University of Dundee) , Saleha Patel (AstraZeneca) , Arjan Snijder (AstraZeneca) , Lisa Wissler (AstraZeneca) , Luc Brunsveld (Technische Universiteit Eindhoven) , Christian Ottmann (Technische Universiteit Eindhoven) , Matthew W. D. Perry (AstraZeneca)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Journal Of Biological Chemistry , VOL 29

State: Published (Approved)
Published: March 2021
Diamond Proposal Number(s): 20016

Abstract: Glucocorticoid receptor (GR) is a ligand-dependent transcription factor that plays a central role in inflammation. GR activity is also modulated via protein–protein interactions, including binding of 14-3-3 proteins induced by GR phosphorylation. However, the specific phosphorylation sites on GR that trigger these interactions and their functional consequences are less clear. Hence, we sought to examine this system in more detail. We used phosphorylated GR peptides, biophysical studies and X-ray crystallography to identify key residues within the ligand binding domain of GR, T524 and S617, whose phosphorylation results in binding of the representative 14-3-3 protein 14-3-3ζ. A kinase screen identified MINK1 as responsible for phosphorylating T524 and ROCK1 for phosphorylating S617; cell-based approaches confirmed the importance of both GR phosphosites and MINK1 but not ROCK1 alone in inducing GR–14-3-3 binding. Together our results provide molecular-level insight into 14-3-3-mediated regulation of GR and highlight both MINK1 and the GR–14-3-3 axis as potential targets for future therapeutic intervention.

Journal Keywords: 14‐3‐3 protein; glucocorticoid receptor; MINK1 kinase; nuclear receptor; phosphorylation; protein‐protein interaction

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)

Other Facilities: ID30B at ESRF

Added On: 28/03/2021 13:42

Documents:
1-s2.0-S002192582100329X-main.pdf

Discipline Tags:

Health & Wellbeing Biochemistry Chemistry Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)