Faculty Research 1990 - 1999

Title

The mitochondrial uncoupling protein gene in brown fat: correlation between DNase I hypersensitivity and expression in transgenic mice.

Document Type

Article

Publication Date

1991

Keywords

Base-Sequence, Blotting-Northern, Brown-Fat: me, Cloning-Molecular, Deoxyribonuclease-I, DNA: ge, Genes-Structural, Genes-Synthetic, Membrane-Proteins: ge, Mice, Mice-Transgenic, Mitochondria: me, Molecular-Sequence-Data, Organ-Specificity, Promoter-Regions-(Genetics), Regulatory-Sequences-Nucleic-Acid, Restriction-Mapping, RNA: ge, RNA-Messenger: ge, SUPPORT-U-S-GOVT-P-H-S

JAX Source

Mol Cell Biol 1991 Aug; 11(8):4147-56.

Grant

NSO8483, HDO8431

Abstract

The mitochondrial uncoupling protein gene is rapidly induced in mouse brown fat following cold exposure. To identify cis-regulatory elements, approximately 50 kb of chromatin surrounding the uncoupling protein gene was examined for its hypersensitivity to DNase I. Seven DNase I-hypersensitive sites were identified in the 5'-flanking DNA, and one site was identified in the 3'-flanking DNA. Transgenic mice with an uncoupling protein minigene were generated by microinjection of fertilized eggs with a transgene containing 3 kb of 5'-flanking DNA and 0.3 kb of 3'-flanking DNA. Expression of the transgene is restricted to brown fat and is cold inducible. Four additional transgenic lines were generated with a second transgene containing a 1.8-kb deletion in the 5'-flanking DNA, and expression of this minigene is absent in all tissues analyzed. A DNase I-hypersensitive site located in the 1.8-kb deletion contains a cyclic AMP response element that binds a brown fat tumor enriched nuclear factor. On the basis of these observations, we propose that a cis-acting regulatory sequence between -3 and -1.2 kb of the 5'-flanking region, possibly at a DNase I-hypersensitive site, is required for controlling uncoupling protein expression in vivo.