Mouse melanocortin-4 receptor gene 5′-flanking region imparts cell specific expression in vitro

Laurence M. Dumont, Chia Shan Wu, Carl J. Aschkenasi, Joel K. Elmquist, Bradford B. Lowell, Kathleen G. Mountjoy

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Weight homeostasis is exquisitely sensitive to changes in the abundance of melanocortin-4 receptor (MC4-R). To begin to understand the factors that regulate MC4-R gene expression, we determined there are no introns in the gene, there are multiple starts of transcription, and a cluster of 3′ ends. A series of MC4-R-luciferase gene reporter chimerics was developed and transfected into cell lines expressing (UMR106; GT1-7; HEK293) and not expressing (Neuro 2A) endogenous MC4-R mRNA. The longest construct, which includes ≈3.3 kb 5′-flanking, 425 bp 5′-untranslated (UTR) and 1852 bp 3′-flanking, significantly increased luciferase reporter gene expression 24-, 13-, and 3-fold compared to pGL3-basic when expressed in HEK293, UMR106, and GT1-7 cells, respectively. Deletion analysis of mMC4-R 5′-flanking cDNA identified full mMC4-R promoter activity within 178 bp upstream of the major start of transcription. The mMC4-R gene structure and reporter chimerics provide a fundamental framework for the identification of specific factors regulating MC4-R gene expression.

Original languageEnglish (US)
Pages (from-to)173-185
Number of pages13
JournalMolecular and Cellular Endocrinology
Volume184
Issue number1-2
DOIs
StatePublished - Nov 26 2001

Fingerprint

Receptor, Melanocortin, Type 4
5' Flanking Region
Genes
Gene expression
Transcription
Luciferases
Reporter Genes
Gene Expression
vpr Genes
Introns
Homeostasis
Complementary DNA
Cells
mouse MC4R protein
In Vitro Techniques
Weights and Measures
Cell Line
Messenger RNA

Keywords

  • Gene structure
  • Melanocortin-4 receptor
  • Promoter activity

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Mouse melanocortin-4 receptor gene 5′-flanking region imparts cell specific expression in vitro. / Dumont, Laurence M.; Wu, Chia Shan; Aschkenasi, Carl J.; Elmquist, Joel K.; Lowell, Bradford B.; Mountjoy, Kathleen G.

In: Molecular and Cellular Endocrinology, Vol. 184, No. 1-2, 26.11.2001, p. 173-185.

Research output: Contribution to journalArticle

Dumont, Laurence M. ; Wu, Chia Shan ; Aschkenasi, Carl J. ; Elmquist, Joel K. ; Lowell, Bradford B. ; Mountjoy, Kathleen G. / Mouse melanocortin-4 receptor gene 5′-flanking region imparts cell specific expression in vitro. In: Molecular and Cellular Endocrinology. 2001 ; Vol. 184, No. 1-2. pp. 173-185.
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