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Cited by 70Estrogen and related hormone effectsReceptor Mechanisms and SignalingAdrenal Hormones and Disorders

Kinetics of glucocorticoid receptor phosphorylation in intact cells. Evidence for hormone-induced hyperphosphorylation after activation and recycling of hyperphosphorylated receptors.

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Ortí, E., Hu, L.M., Munck, A.·1993·Journal of Biological Chemistry
DOI 10.1016/s0021-9258(18)53026-x
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Abstract

Glucocorticoid receptors are basally phosphorylated in the absence of hormone and become hyperphosphorylated after hormone treatment of intact cells. To determine the sequence of changes which the receptor undergoes following hormone binding, we analyzed the kinetics of receptor phosphorylation in WEHI-7 mouse thymoma cells and in stably transfected Chinese hamster ovary cells that overexpress the mouse receptor. No major differences were found between these two cell types. Cells were preincubated with 32P(i) and [35S] methionine to label the receptors metabolically. The phosphate content of the receptor protein was determined from the ratio of 32P to 35S in radioactive gel slices after immunopurification and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Hormone-induced increases in phosphorylation were seen as early as 5 min after adding hormone and persisted for 20 h. Analysis of newly formed cytosolic and nuclear-bound activated (DNA-binding) receptors showed that activation precedes hyperphosphorylation. Nonactivated receptors, both unliganded and hormone-liganded, also became hyperphosphorylated but more slowly than activated receptors. The rate of receptor dephosphorylation, determined by chasing with unlabeled P(i), was much slower than the rate of phosphorylation or of hormone dissociation and appeared to be slightly increased by agonists and by the antagonist RU486 (which does not cause hyperphosphorylation). Mutant WEHI-7 cells lacking cAMP-dependent protein kinase activity gave basal and hormone-induced receptor phosphorylation indistinguishable from wild type cells. We conclude that (a) the substrate for hormone-dependent hyperphosphorylation is the activated hormone-receptor complex; (b) most hyperphosphorylated receptors are recycled and reutilized in hyperphosphorylated form; (c) control of receptor phosphorylation may not be cell-specific; (d) cAMP-dependent protein kinase is not involved directly or indirectly in phosphorylating major sites on the receptor in vivo.

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  • Multiple kinases differentially phosphorylate the GR at serine residues S203, S211 and S226, regulating GR nuclear translocation and GR-dependent gene transcription.

    “Multiple kinases have been reported to differentially phosphorylate the GR at its serine residues, S203, S211 and S226, an effect which regulates GR nuclear translocation and GR-dependent gene transcription.25, 26, 27, 28, 29”

    From Antidepressants increase human hippocampal neurogenesis by activating the glucocorticoid receptor (2011) · cites this paper as [28]

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Authors
Ortí, E., Hu, L. M., & Munck, A.
ORCID
Hu, L.M.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1993-04
Volume
268
Issue
11
Pages
7779-7784
ISSN
0021-9258 (ISSN-L)
DOI
10.1016/s0021-9258(18)53026-x
OpenAlex
W1479704824
Cited by
70
References
20
Citations by year
2018: 3 · 2017: 1 · 2016: 1 · 2013: 2 · 2012: 1
FWCI
4.01
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (1f05tw003923-01, 5r01dk003535-39)
  • National Cancer Institute (CA 23 108, P30CA023108)
  • Fogarty International Center (1F05 TW03923-01)
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK 03535)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
Keywords
glucocorticoid receptor, receptor phosphorylation, hyperphosphorylation, receptor recycling, protein kinase A, RU486
MeSH terms
Animals, Cell Nucleus, Cytosol, Cricetinae, Kinetics, Methionine, Mutation, Phosphates, Phosphoproteins, Phosphorus Radioisotopes, Phosphorylation, Protein Kinases, Receptors, Glucocorticoid, Sulfur Radioisotopes, Thymoma, Thymus Neoplasms, Time Factors, Transfection, Triamcinolone Acetonide, Tumor Cells, Cultured, Mifepristone, CHO Cells, Mice

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