A sequence element in the GLUT4 gene that mediates repression by insulin

David W Cooke, M. Daniel Lane

Research output: Contribution to journalArticle

Abstract

Prolonged treatment of 3T3-L1 adipocytes decreases expression of GLUT4, the insulin-responsive glucose transporter. Expression of promoter-reporter gene constructs that contained 2900 or 785 base pairs of 5'-flanking region of the murine GLUT4 gene was down-regulated by insulin (p <0.0005), whereas expression of constructs that contained 641, 469, or 78 base pairs of 5'- flanking region was not. Nuclear extract from 3T3-L1 adipocytes protected the region from -707 to -681 in the GLUT4 5'-flanking region from DNase I digestion. Using an oligonucleotide probe that corresponded to this footprinted region, two major protein-DNA complexes were identified by a gel mobility shift assay. Southwestern analysis identified four protein bands with molecular masses from 38 to 46 kDa that bound to the insulin-responsive region probe. A reporter gene construct in which bases -706 to -676 were deleted was not repressed by insulin treatment, confirming that this sequence is necessary for the repression of the GLUT4 promoter by insulin in 3T3-L1 adipocytes. This sequence does not show homology to previously described insulin response elements and thus represents a distinct mechanism of gene regulation by insulin.

Original languageEnglish (US)
Pages (from-to)6210-6217
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number11
DOIs
StatePublished - Mar 13 1998

Fingerprint

Genes
Insulin
5' Flanking Region
Adipocytes
Reporter Genes
Base Pairing
Glucose Transporter Type 4
Oligonucleotide Probes
Deoxyribonuclease I
Molecular mass
Response Elements
Electrophoretic Mobility Shift Assay
Gene expression
Digestion
Assays
Proteins
Gels
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

A sequence element in the GLUT4 gene that mediates repression by insulin. / Cooke, David W; Lane, M. Daniel.

In: Journal of Biological Chemistry, Vol. 273, No. 11, 13.03.1998, p. 6210-6217.

Research output: Contribution to journalArticle

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