Selective regulation of Mmp13 by 1,25(OH)2D3, PTH, and Osterix through distal enhancers.
Matrix metalloproteinase 13 (MMP13, collagenase-3) is a vital component for chondrocyte and osteoblast maturation, and is aberrantly expressed in numerous disease states. At the transcriptional level, Mmp13 is controlled by many different growth factors and hormones. Most notably, Mmp13 is regulated by the vitamin D hormone (1,25(OH)2D3), parathyroid hormone (PTH), and several cytokines. These activities occur through participation by the transcription factors VDR, RUNX2, FOS, JUN, and Osterix (OSX), respectively. Recently, we discovered that Mmp13 is regulated by elements quite distal to the transcriptional start site -10, -20, and -30kb upstream. These enhancers, along with minor contributions from the region proximal to the promoter, are responsible for the ligand inducible and, most strikingly, the basal activities of Mmp13 gene regulation. Here, we found that the actions of PTH and OSX do not occur through the -10kb VDR bound enhancer. Rather, the -30kb RUNX2 bound enhancer and the promoter proximal regions were essential for activity. Through RUNX2 deletion and OSX overexpression in cells, we showed a specific role for OSX in Mmp13 regulation. Finally, we created an in vivo CRISPR deleted -10kb enhancer mouse model. Despite normal bone density and growth, they fail to up-regulate Mmp13 in response to 1,25(OH)2D3. These data are consistent with those obtained through UAMS osteoblast cell culture and further define the specific roles of distal enhancers in the regulation of Mmp13.
University of Wisconsin at Madison, Madison, WI 53706, USA. Electronic address: mmeyer@biochem.wisc.edu.
Mark B
MB
Meyer
University of Wisconsin at Madison, Madison, WI 53706, USA.
Nancy A
NA
Benkusky
University of Wisconsin at Madison, Madison, WI 53706, USA.
Melda
M
Onal
University of Wisconsin at Madison, Madison, WI 53706, USA.
J Wesley
JW
Pike
D000818:Animals; D002117:Calcitriol / Q000494:pharmacology*; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005260:Female; D000072669:Gene Editing; D005786:Gene Expression Regulation; D008297:Male; D053509:Matrix Metalloproteinase 13 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010281:Parathyroid Hormone / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D018167:Receptors, Calcitriol / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D050659:Core Binding Factor Alpha 1 Subunit; D010281:Parathyroid Hormone; D018167:Receptors, Calcitriol; C493716:Runx2 protein, mouse; D000076182:Sp7 Transcription Factor; C479466:Sp7 protein, mouse; D014157:Transcription Factors; D053509:Matrix Metalloproteinase 13; C507081:Mmp13 protein, mouse; D002117:Calcitriol
1,25(OH)(2)D(3); ChIP-seq; Histone modification; Mmp13; Osterix; PTH; RUNX2; Sp7; VDR; Vitamin D