Identification of glucose-responsive and insulin-responsive elements in promoter of mouse ob gene

Fang Nian Wang, Chun Gu Ma, Yan Ling Zhang, Nai Xian Zhang, You Mei Chen, Qi Qun Tang, Hou Yan Song

Research output: Contribution to journalArticlepeer-review

Abstract

Glucose and insulin stimulate leptin gene expression in vitro and in vivo. To identify cis-elements that are responsible for the glucose and insulin effects, mouse 3T3-L1 adipocytes were transiently transfected with reporter constructs with serial deletions in mouse ob gene promoter. The cis-elements were identified with gel mobility shift assays (GMSA), DNase I footprint assays and PCR mediated site-directed mutation assays. Transient transfections detected a negative cis-acting element, a glucose-responsive element (GLRE), and an insulin-responsive element (IRE) in the region from -1 719 bp to -1 452 bp of mouse ob gene. This region does not contain any known GLRE or IRE. GMSA identified a DNA binding protein which specifically binds a native probe prepared from mouse ob gene promoter ( -1 719 bp to -1 452 bp), and the binding was repressed by glucose or insulin. DNase I footprint assays and PCR mediated site-directed mutations assays identified that the binding motif AGCAAAA, spanning -1 698 bp to -1 692 bp of the mouse ob gene promoter, was responsible for the effects of glucose and insulin on ob gene expression. These studies suggest that a negative cis-acting element is located between - 1 719 bp and - 1 452 bp of the mouse ob gene promoter, and glucose and insulin simulate mouse ob gene expression by repressing the binding of a transcription factor to this element. This element, AGCAAAA, spanning - 1 698 bp to - 1 692 bp is a novel GLRE and IRE.

Original languageEnglish (US)
Pages (from-to)543-544
Number of pages2
JournalActa Biochimica et Biophysica Sinica
Volume32
Issue number5
StatePublished - 2000
Externally publishedYes

Keywords

  • Glucose
  • Insulin
  • Leptin
  • Ob gene
  • Response element

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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