p38 Mitogen-activated protein kinase (MAPK) is a key mediator in glucocorticoid-induced apoptosis of lymphoid cells: correlation between p38 MAPK activation and site-specific phosphorylation of the human glucocorticoid receptor at serine 211.
Glucocorticoids (GCs) induce apoptosis in lymphoid cells through activation of the GC receptor (GR). We have evaluated the role of p38, a MAPK, in lymphoid cell apoptosis upon treatment with the synthetic GCs dexamethasone (Dex) or deacylcortivazol (DAC). The highly conserved phosphoprotein p38 MAPK is activated by specific phosphorylation of its threonine180 and tyrosine182 residues. We show that Dex and DAC stimulate p38 MAPK phosphorylation and increase the mRNA of MAPK kinase 3, a specific immediate upstream activator of p38 MAPK. Enzymatic assays confirmed elevated activity of p38 MAPK. Pharmacological inhibition of p38 MAPK activity was protective against GC-driven apoptosis in human and mouse lymphoid cells. In contrast, inhibition of the MAPKs, ERK and cJun N-terminal kinase, enhanced apoptosis. Activated p38 MAPK phosphorylates specific downstream targets. Because phosphorylation of the GR is affected by MAPKs, we examined its phosphorylation state in our system. We found serine 211 of the human GR to be a substrate for p38 MAPK both in vitro and intracellularly. Mutation of this site to alanine greatly diminished GR-driven gene transcription and apoptosis. Our results clearly demonstrate a role for p38 MAPK signaling in the pathway of GC-induced apoptosis of lymphoid cells.
Miller
Aaron L
AL
Department of Human Biological Chemistry and Genetics, The University of Texas Medical Branch, 301 University Boulevard, Room 5.104, Medical Research Building, Route 1068, Galveston, Texas 77555-1068, USA.
Webb
M Scott
MS
Copik
Alicja J
AJ
Wang
Yongxin
Y
Johnson
Betty H
BH
Kumar
Raj
R
Thompson
E Brad
EB
D000409:Alanine / Q000737:chemistry; D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival; D004247:DNA / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D004789:Enzyme Activation; D005434:Flow Cytometry; D005938:Glucocorticoids / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D008214:Lymphocytes / Q000473:pathology*; D048371:MAP Kinase Kinase 3 / Q000378:metabolism; D020935:MAP Kinase Signaling System; D051379:Mice; D008954:Models, Biological; D009154:Mutation; D010766:Phosphorylation; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000378:metabolism; D011419:Propidium / Q000494:pharmacology; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000737:chemistry*; D012694:Serine / Q000737:chemistry; D015398:Signal Transduction; D013912:Threonine / Q000737:chemistry; D013997:Time Factors; D015533:Transcriptional Activation; D014162:Transfection; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D005938:Glucocorticoids; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D013912:Threonine; D011419:Propidium; D012694:Serine; D003907:Dexamethasone; D004247:DNA; D048051:p38 Mitogen-Activated Protein Kinases; D048371:MAP Kinase Kinase 3; D000409:Alanine