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

被引:19
|
作者
Dumont, LM
Wu, CS
Aschkenasi, CJ
Elmquist, JK
Lowell, BB
Mountjoy, KG
机构
[1] Univ Auckland, Fac Med & Hlth Sci, Div Phys, Res Ctr Dev Med & Biol, Auckland 1, New Zealand
[2] Univ Auckland, Fac Med & Hlth Sci, Div Mol Med, Auckland 1, New Zealand
[3] Harvard Univ, Beth Israel Deaconess Med Ctr, Div Endocrinol, Boston, MA 02215 USA
基金
英国惠康基金;
关键词
melanocortin-4; receptor; gene structure; promoter activity;
D O I
10.1016/S0303-7207(01)00558-5
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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 approximate to3.3 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. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.
引用
收藏
页码:173 / 185
页数:13
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