Glucocorticoids stimulate human sgk1 gene expression by activation of a GRE in its 5'-flanking region

Omar A Itani; Kang Z Liu; Kristyn L Cornish; Jason R Campbell; Christie P Thomas

doi:10.1152/ajpendo.00021.2002

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Glucocorticoids stimulate human sgk1 gene expression by activation of a GRE in its 5'-flanking region

Journal article

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Glucocorticoids stimulate human sgk1 gene expression by activation of a GRE in its 5'-flanking region

Omar A Itani, Kang Z Liu, Kristyn L Cornish, Jason R Campbell and Christie P Thomas

American journal of physiology: endocrinology and metabolism, Vol.283(5), pp.E971-E979

11/2002

DOI: 10.1152/ajpendo.00021.2002

PMID: 12376324

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Abstract

In lung and collecting duct epithelia, glucocorticoid (GC)-stimulated Na+ transport is preceded by an increase in the protein kinase sgk1, which in turn regulates the activity of the epithelial Na+ channel (ENaC). We investigated the mechanism for GC-regulated human sgk1 expression in lung and renal epithelia. sgk1 mRNA was increased in these epithelia by GCs, and this was inhibited by actinomycin D and superinduced by cycloheximide, consistent with a transcriptional effect that did not require protein synthesis. To understand the basis for transcriptional regulation, the transcription initiation site was mapped and the 5'-flanking region cloned by PCR. A 3-kb fragment of the upstream region was coupled to luciferase and transfected into A549 cells. By deletion analysis, an imperfect GC response element (GRE) was identified that was necessary and sufficient for GC responsiveness. When tested with cell extracts, a specific protein recognized by an anti-GC receptor (GR) antibody bound the GRE in gel mobility shift assays. We conclude that GCs stimulate sgk1 expression in human epithelial cells via activation of a GRE in the 5'-flanking region of sgk1.

Amino Acid Sequence

Gene Expression Regulation, Enzymologic - drug effects

Respiratory Mucosa - cytology

Humans

Cells, Cultured

Protein-Serine-Threonine Kinases - genetics

Molecular Sequence Data

RNA, Messenger - analysis

Receptors, Glucocorticoid - metabolism

Sodium - metabolism

5' Flanking Region - genetics

Nuclear Proteins

Immediate-Early Proteins

Epithelial Sodium Channels

Transcription, Genetic - physiology

Dexamethasone - pharmacology

Receptors, Glucocorticoid - genetics

Sodium Channels - metabolism

Glucocorticoids - pharmacology

Respiratory Mucosa - physiology

Electrophoretic Mobility Shift Assay

Enhancer Elements, Genetic - genetics

Details

Title: Subtitle: Glucocorticoids stimulate human sgk1 gene expression by activation of a GRE in its 5'-flanking region
Creators: Omar A Itani - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa 52246, USA
Kang Z Liu
Kristyn L Cornish
Jason R Campbell
Christie P Thomas
Resource Type: Journal article
Publication Details: American journal of physiology: endocrinology and metabolism, Vol.283(5), pp.E971-E979
DOI: 10.1152/ajpendo.00021.2002
PMID: 12376324
NLM abbreviation: Am J Physiol Endocrinol Metab
ISSN: 0193-1849
eISSN: 1522-1555
Publisher: United States
Grant note: R01 DK054348 / NIDDK NIH HHS DK-54348 / NIDDK NIH HHS
Language: English
Date published: 11/2002
Academic Unit: Stead Family Department of Pediatrics; Obstetrics and Gynecology; Nephrology; Internal Medicine
Record Identifier: 9983986367802771

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