Open AccessReconstitution of the vitamin D-responsive osteocalcin transcription unit in Saccharomyces cerevisiae.
Author(s) -
Donald P. McDonnell,
J. Wesley Pike,
David J. Drutz,
Tauseef R. Butt,
O’Malley Bw
Publication year - 1989
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.9.8.3517
Subject(s) - biology , enhancer , saccharomyces cerevisiae , osteocalcin , reporter gene , gene , transcription factor , transcription (linguistics) , promoter , regulation of gene expression , microbiology and biotechnology , calcitriol receptor , yeast , gene expression , genetics , biochemistry , alkaline phosphatase , linguistics , philosophy , enzyme
The human osteocalcin gene is regulated in mammalian osteoblasts by 1,25(OH)2D3-dependent and -independent mechanisms. The sequences responsible for this activity have been mapped to within the -1339 region of the gene. We show here that this enhancer region functions analogously in Saccharomyces cerevisiae cells engineered to produce active 1,25(OH)2D3 receptor. When fused to the proximal promoter elements of the yeast iso-1-cytochrome c gene, the enhancer demonstrated substantial promoter activity. This activity was elevated further by 1,25(OH)2D3 when the reporter constructs were assayed in cells containing the 1,25(OH)2D3 receptor. This system affords a model for 1,25(OH)2D3 action and represents a simple assay system that will enable definition of the important cis-acting regulatory sequences within the osteocalcin gene and identification of their cognate transcription factors.