Characterization of the mouse tartrate-resistant acid phosphatase (trap) gene promoter

Abstract

Tartrate-resistant acid phosphatase (TRAP) is an iron-binding protein that is highly expressed in osteoclasts. To characterize the regulation of TRAP gene expression, progressive 5' and 3' deletions of a 1.8 kb fragment containing the 5'-flanking sequence were fused to a luciferase reporter gene. Two nonoverlapping regions of this 1.8 kb fragment had promoter activity. The upstream promoter (P1) was located within the region from -881 bp to -463 bp relative to the ATG, while the downstream promoter (P2) was located between -363 bp to -1 bp in a region we have previously shown to be an intron in transcripts originating from the upstream promoter. A putative repressor region for the P2 promoter at -1846 bp to -1240 bp and a putative enhancer region at -962 bp to -881 bp relative to the ATG were identified. PCR analysis of promoter-specific transcription of the TRAP gene in various murine tissues showed that both promoters were active in several tissues. Transferrin-bound iron increased P1 promoter activity 2.5-fold and hemin decreased P1 promoter activity, but neither had any effect on P2 activity. These data show that the transcriptional regulation of the TRAP gene is complex and that iron may play a key role in TRAP gene regulation.

Hexane-Soluble Fraction of the Common Fig, Ficus carica, Inhibits Osteoclast Differentiation in Murine Bone Marrow-Derived Macrophages and RAW 264.7 Cells

“…Genes such as TRAP, cathepsin K, and MMP-9, which are specifically induced during terminal osteoclast differentiation, contain multiple NFAT and AP-1 binding sites (Reddy et al, 1995;Motyckova et al, 2001). Both AP-1 and NF-κB binding sites are present within the promoter region of the NFATc1 gene (Zhou et al, 2002).…”

Calcium/calmodulin-signaling supports TRPV4 activation in osteoclasts and regulates bone mass

“…(17) The pcDNA3.1 plasmidfree AccI-digested inserts of the Trpv4 transgenes containing the Trap gene promoter and bovine growth hormone polyadenylation (BGH polyA) signal sequences were injected into fertilized ova of B6C3F1 mice. C57BL/6 mice were then used for generating the osteoclast-specific Trpv4 transgenic lines.…”

Inactivation of Glycogen Synthase Kinase-3β Is Required for Osteoclast Differentiation

“…Generation of Transgenic Mice-The constitutively active GSK-3␤ (GSK3␤-S9A) mutant cDNA was fused to the mouse TRAP gene promoter as described previously (29,30). For generating transgenic mice, we used the standard pronuclear injection method with C57BL/6 mice (The Jackson Laboratory).…”