Hormone-induced hyperphosphorylation of specific phosphorylated sites in the mouse glucocorticoid receptor

Bodwell, Jack E.; Hu, Jiong-Ming; Ortí, Eduardo; Munck, Allan

doi:10.1016/0960-0760(94)00157-h

Cited by 58 publications

(60 citation statements)

References 13 publications

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“…Based on these general phenomena, we hypothesize that posttranslational modification of the GR is important in regulating GC signaling within cells. The human GR is known to be a phospho-protein; however, previous studies have not thoroughly investigated the role of phosphorylation in GR signaling (5). Therefore, we initially utilized mass spectrometry to identify additional residues within the human GR that were phosphorylated in response to the synthetic GC, Dex.…”

Section: Resultsmentioning

confidence: 99%

Glycogen Synthase Kinase 3β-Mediated Serine Phosphorylation of the Human Glucocorticoid Receptor Redirects Gene Expression Profiles

Galliher-Beckley¹,

Williams²,

Collins

et al. 2008

Molecular and Cellular Biology

114

102

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Aberrant glycogen synthase kinase 3␤ (GSK-3␤) activity is associated with the progression of several pathological conditions such as diabetes, Alzheimer's, and cancer. GSK-3␤ regulates cellular processes by directly phosphorylating metabolic enzymes and transcription factors. Here, we discovered a new target for GSK-3␤ phosphorylation: the human glucocorticoid receptor (GR). Glucocorticoid signaling is essential for life and regulates diverse biological functions from cell growth to metabolism to apoptosis. Specifically, we found hormone-dependent GR phosphorylation on serine 404 by GSK-3␤. Cells expressing a GR that is incapable of GSK-3␤ phosphorylation had a redirection of the global transcriptional response to hormone, including the activation of additional signaling pathways, in part due to the altered ability of unphosphorylatable GR to recruit transcriptional cofactors CBP/p300 and the p65 (RelA) subunit of NF-B. Furthermore, GSK-3␤-mediated GR phosphorylation inhibited glucocorticoid-dependent NF-B transrepression and attenuated the glucocorticoid-dependent cell death of osteoblasts. Collectively, our results describe a novel convergence point of the GSK-3␤ and the GR pathways, resulting in altered hormone-regulated signaling. Our results also provide a mechanism by which GSK-3␤ activity can dictate how cells will ultimately respond to glucocorticoids.

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Section: Resultsmentioning

confidence: 99%

Glycogen Synthase Kinase 3β-Mediated Serine Phosphorylation of the Human Glucocorticoid Receptor Redirects Gene Expression Profiles

Galliher-Beckley¹,

Williams²,

Collins

et al. 2008

Molecular and Cellular Biology

114

102

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“…Retention at the nuclear matrix could also be reversed by a slow reversal of ligand-induced posttranslational modification. We note that GR is a phosphoprotein and that the phosphorylation of GR following treatment with hormone agonist is only slowly reversed upon hormone withdrawal (4,38,44,65). Further, GRs are differentially phosphorylated following treatment with RU486 (42), which appears to lead to the permanent localization of GR to the nucleus following hormone withdrawal (36,78,83).…”

Section: Discussionmentioning

confidence: 99%

Discrimination between NL1- and NL2-Mediated Nuclear Localization of the Glucocorticoid Receptor

Savory

Hsu

Laquian

et al. 1999

Molecular and Cellular Biology

202

182

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Glucocorticoid receptor (GR) cycles between a free liganded form that is localized to the nucleus and a heat shock protein (hsp)-immunophilin-complexed, unliganded form that is usually localized to the cytoplasm but that can also be nuclear. In addition, rapid nucleocytoplasmic exchange or shuttling of the receptor underlies its localization. Nuclear import of liganded GR is mediated through a well-characterized sequence, NL1, adjacent to the receptor DNA binding domain and a second, uncharacterized motif, NL2, that overlaps with the ligand binding domain. In this study we report that rapid nuclear import (half-life [t 1/2 ] of 4 to 6 min) of agonist-and antagonist-treated GR and the localization of unliganded, hsp-associated GRs to the nucleus in G 0 are mediated through NL1 and correlate with the binding of GR to pendulin/importin ␣. By contrast, NL2-mediated nuclear transfer of GR occurred more slowly (t 1/2 ‫؍‬ 45 min to 1 h), was agonist specific, and appeared to be independent of binding to importin ␣. Together, these results suggest that NL2 mediates the nuclear import of GR through an alternative nuclear import pathway. Nuclear export of GR was inhibited by leptomycin B, suggesting that the transfer of GR to the cytoplasm is mediated through the CRM1-dependent pathway. Inhibition of GR nuclear export by leptomycin B enhanced the nuclear localization of both unliganded, wild-type GR and hormone-treated NL1 ؊ GR. These results highlight that the subcellular localization of both liganded and unliganded GRs is determined, at least in part, by a flexible equilibrium between the rates of nuclear import and export.The predominant pathway for the nuclear import of transcription factors and other nuclear regulatory proteins originates with the interaction of importin ␣-like proteins (also called karyopherin ␣, Rch1/hSRP␣, hSRP1/NPI-1, and pendulin/OHO31) with specific nuclear localization sequences (NLSs), which contain closely spaced arrangements of five to eight basic amino acids (31,62,64). For DNA sequence-specific transcription factors, NLSs generally colocalize with their DNA binding domains (DBDs), which appears to reflect a coevolutionary selective pressure to ensure that proteins that bind DNA are able to access the nucleus (52). Nuclear export, by contrast, occurs through alternative pathways, which for many proteins involves the binding of CRM1 (exportin 1) to hydrophobic nuclear export sequences (26,90).However, some transcription factors, including the glucocorticoid hormone receptor (GR), contain additional NLSs that occur in other regions of the proteins (69,89,95,99). In at least some instances, the presence of these additional NLSs has been found to reflect a requirement for specialized or tightly regulated nuclear localization of the protein. For example, the nuclear localization potential of one of the two NLSs in the adenovirus E1A protein is active only during early development (92), while two of the three c-abl NLSs promote nuclear localization of c-abl only in certain cell types (97, 99). T...

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“…GR is a phosphoprotein, 49 for which the role of phosphorylation is poorly understood. Several reports have implied this mechanism in regulating glucocorticoid responses.…”

Section: Discussionmentioning

confidence: 99%

Glucocorticoid receptor down-Regulates c-jun amino terminal kinases induced by tumor necrosis factor ? in fetal rat hepatocyte primary cultures

et al. 1999

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The effect of dexamethasone on Jun N-terminal kinase (JNK) activity was assayed by using fetal hepatocytes in primary culture. The addition of tumor necrosis factor ␣ (TNF-␣) caused an increase in JNK in a dose-and timedependent manner. We show that activation of JNK by this extracellular signal is inhibited by dexamethasone in a dose-dependent fashion. This inhibitory effect was observed in cells treated for 10 minutes with dexamethasone in the presence of protein phosphatase inhibitors such as orthovanadate or okadaic acid, or in cells previously treated with actinomycin D. Glucocorticoid receptor (GR) can be precipitated with the fusion protein, GST-c-Jun (1-79), bound to agarose beads. However, the inhibitory effect of glucocorticoids on JNK activity was also observed using ATF-2 as substrate. In addition, dexamethasone inhibits JNK phosphorylation induced by TNF-␣. Finally, we show that GR can also be phosphorylated in tyrosine residues in response to TNF-␣ and epidermal growth factor (EGF) upon ligand-binding. Our results suggest that the antiinflammatory effect of glucocorticoids on the inflammatory pathways induced by TNF-␣ can be explained, at least in part, by modulating JNK activity through a direct proteinprotein interaction; the JNK phosphorylation and tyrosinephosphorylation state of GR may be regulatory steps also involved in that effect. (HEPATOLOGY 1999;29:849-857.)

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Hormone-induced hyperphosphorylation of specific phosphorylated sites in the mouse glucocorticoid receptor

Cited by 58 publications

References 13 publications

Glycogen Synthase Kinase 3β-Mediated Serine Phosphorylation of the Human Glucocorticoid Receptor Redirects Gene Expression Profiles

Glycogen Synthase Kinase 3β-Mediated Serine Phosphorylation of the Human Glucocorticoid Receptor Redirects Gene Expression Profiles

Discrimination between NL1- and NL2-Mediated Nuclear Localization of the Glucocorticoid Receptor

Glucocorticoid receptor down-Regulates c-jun amino terminal kinases induced by tumor necrosis factor ? in fetal rat hepatocyte primary cultures

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