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PMID: 15879558 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The glucocorticoid receptor blocks P-TEFb recruitment by NFkappaB to effect promoter-specific transcriptional repression.

Genes & development ·Vol. 19 ·No. 9 ·2005-05-01 ·Pages 1116-27

Luecke HF, Yamamoto KR

Abstract

To investigate the determinants of promoter-specific gene regulation by the glucocorticoid receptor (GR), we compared the composition and function of regulatory complexes at two NFkappaB-responsive genes that are differentially regulated by GR. Transcription of the IL-8 and IkappaBalpha genes is stimulated by TNFalpha in A549 cells, but GR selectively represses IL-8 mRNA synthesis by inhibiting Ser2 phosphorylation of the RNA polymerase II (pol II) C-terminal domain (CTD). The proximal kappaB elements at these genes differ in sequence by a single base pair, and both recruited RelA and p50. Surprisingly, GR was recruited to both of these elements, despite the fact that GR failed to repress the IkappaBalpha promoter. Rather, the regulatory complexes formed at IL-8 and IkappaBalpha were distinguished by differential recruitment of the Ser2 CTD kinase, P-TEFb. Disruption of P-TEFb function by the Cdk-inhibitor, DRB, or by small interfering RNA selectively blocked TNFalpha stimulation of IL-8 mRNA production. GR competed with P-TEFb recruitment to the IL-8 promoter. Strikingly, IL-8 mRNA synthesis was repressed by GR at a post-initiation step, demonstrating that promoter proximal regulatory sequences assemble complexes that impact early and late stages of mRNA synthesis. Thus, GR accomplishes selective repression by targeting promoter-specific components of NFkappaB regulatory complexes.

MeSH Terms
Chromatin Immunoprecipitation Dichlororibofuranosylbenzimidazole/pharmacology Gene Expression Regulation Glutathione Transferase/metabolism Humans I-kappa B Proteins/genetics Interleukin-8/genetics Models, Biological NF-KappaB Inhibitor alpha NF-kappa B/metabolism Oligonucleotides Phosphorylation Positive Transcriptional Elongation Factor B/antagonists & inhibitors,metabolism Promoter Regions, Genetic/genetics RNA Polymerase II/antagonists & inhibitors,metabolism RNA, Small Interfering/pharmacology Receptors, Glucocorticoid/metabolism Reverse Transcriptase Polymerase Chain Reaction Transcription, Genetic/drug effects Tumor Cells, Cultured Tumor Necrosis Factor-alpha/metabolism
Chemicals
I-kappa B Proteins Interleukin-8 NF-kappa B NFKBIA protein, human Oligonucleotides RNA, Small Interfering Receptors, Glucocorticoid Tumor Necrosis Factor-alpha NF-KappaB Inhibitor alpha Dichlororibofuranosylbenzimidazole Glutathione Transferase Positive Transcriptional Elongation Factor B RNA Polymerase II
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Luecke Hans F
Department of Cellular and Molecular Pharmacology, University of California-San Francisco, San Francisco, CA 94107-2280, USA.
Yamamoto Keith R
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1297105
Published
2005-05-01
Pages
1116-27
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC1091745
Subset
IM
Grants
NCI NIH HHS · R01 CA020535 · United States
NCI NIH HHS · R01 CA 20535 · United States
External Links
PubMed source DOI: 10.1101/gad.1297105 PMC Full Text View journal details
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