Regulation by estrogen through the 5'-flanking region of the mouse calbindin-D28k gene

Mol Endocrinol. 1995 Mar;9(3):319-26. doi: 10.1210/mend.9.3.7776978.


Mouse calbindin-D28k expression is regulated in vivo by estradiol in ovaries, uterus, and oviduct. To determine whether estrogen can have an effect on the transcription of the calbindin-D28k gene, the human breast cancer cells T47D were transiently transfected with a plasmid containing a 1.1 kilobase (kb) PstI/SacII fragment (-1075/+34) of the mouse calbindin gene ligated to the chloramphenicol acetyltransferase (CAT) gene and cotransfected with human estrogen receptor expression vector. T47D cells, transfected and treated with estradiol (10(-11) - 10(-7) M for 64-65 h), exhibited a dose-dependent increase in CAT activity (up to 6.2-fold). Transfection of MCF-7 breast cancer cells with the chimeric gene construct alone also resulted in an estradiol-dependent induction in CAT activity. Deletion mutant analysis demonstrated that there are two regions of the mouse calbindin-D28k promoter (between -1075/-702 and between -175/-78) that contribute to the induction by estradiol. These fragments, when linked to the thymidine kinase promoter to construct a heterologous promoter chimera, were able to convert the thymidine kinase promoter to estrogen responsiveness. In these regions there are multiple imperfect half-palindromic estrogen-responsive elements. Gel retardation assays demonstrated weak protein-DNA interactions that were competed with cold oligonucleotide containing the vitellogenin estrogen-response element. These findings indicate that the mouse calbindin-D28k promoter is capable of conferring estrogen responsiveness, which may be mediated by several imperfect half-palindromic estrogen-responsive elements, and suggest, in light of previous studies concerning 1,25-dihydroxyvitamin D regulation, multiple steroid regulation of the calbindin-D28k gene.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Breast Neoplasms
  • Calbindin 1
  • Calbindins
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Estradiol / pharmacology*
  • Female
  • Gene Expression Regulation / drug effects*
  • Humans
  • Mice
  • Molecular Sequence Data
  • Oligonucleotides / metabolism
  • Plasmids
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins
  • Repetitive Sequences, Nucleic Acid
  • S100 Calcium Binding Protein G / genetics*
  • Thymidine Kinase / genetics
  • Transcription, Genetic / drug effects
  • Transfection
  • Tumor Cells, Cultured


  • CALB1 protein, human
  • Calb1 protein, mouse
  • Calbindin 1
  • Calbindins
  • Oligonucleotides
  • Recombinant Fusion Proteins
  • S100 Calcium Binding Protein G
  • Estradiol
  • Chloramphenicol O-Acetyltransferase
  • Thymidine Kinase