Organization and expression of the mouse spot35/calbindin-D28k gene: identification of the vitamin D-responsive promoter region.
Spot35/calbindin-D28k is transcriptionally induced by 1,25-dihydroxyvitamin D3 in the intestine and kidney but not the brain. The structural organization of the mouse calbindin-D28k gene has now been determined and its transcriptional activation examined. Two direct repeat motifs homologous to other vitamin D-responsive elements were identified at positions -2295 to -2281 and -1160 to -1146. Undirectional deletion analyses of the 5' flanking region indicated that the sequence between -1339 and -1038 conferred vitamin D sensitivity on a reporter construct. Furthermore, the proximal (AGGTGAtgaAAGTCA), but not the distal (AGGTGAacaGGTTCA), direct repeat enhanced 1,25(OH)2D3-stimulated transcription in an orientation-dependent manner when placed upstream of the heterologous SV40 promoter. In conclusion, the proximal direct repeat plays a critical role in mediating the vitamin D-induced expression of calbindin-D28k in the kidney.
Takeda
T
T
Research Laboratory for Molecular Genetics, Niigata University, Japan.
Arakawa
M
M
Kuwano
R
R
D000818:Animals; D001483:Base Sequence; D064092:Calbindin 1; D064026:Calbindins; D002117:Calcitriol / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000235:genetics*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004285:Dogs; D005091:Exons; D007438:Introns; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D064030:S100 Calcium Binding Protein G / Q000235:genetics*; D013552:Swine; D014807:Vitamin D / Q000494:pharmacology*
C576499:Calb1 protein, mouse; D064092:Calbindin 1; D064026:Calbindins; D002135:Calcium-Binding Proteins; D009419:Nerve Tissue Proteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D064030:S100 Calcium Binding Protein G; D014807:Vitamin D; D002117:Calcitriol