TY - JOUR
T1 - Mouse melanocortin-4 receptor gene 5′-flanking region imparts cell specific expression in vitro
AU - Dumont, Laurence M.
AU - Wu, Chia Shan
AU - Aschkenasi, Carl J.
AU - Elmquist, Joel K.
AU - Lowell, Bradford B.
AU - Mountjoy, Kathleen G.
N1 - Funding Information:
We thank Professor P. Mellon for generously providing the GT1-7 cells. This work was supported by The Wellcome Trust, The Health Research Council of NZ, and The NZ Lottery Board.
PY - 2001/11/26
Y1 - 2001/11/26
N2 - 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.
AB - 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.
KW - Gene structure
KW - Melanocortin-4 receptor
KW - Promoter activity
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U2 - 10.1016/S0303-7207(01)00558-5
DO - 10.1016/S0303-7207(01)00558-5
M3 - Article
C2 - 11694353
AN - SCOPUS:0035955871
SN - 0303-7207
VL - 184
SP - 173
EP - 185
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
IS - 1-2
ER -