Runx2 recruits p300 to mediate parathyroid hormone's effects on histone acetylation and transcriptional activation of the matrix metalloproteinase-13 gene.
PTH regulates transcription of a number of genes involved in bone remodeling and calcium homeostasis. We have previously shown that the matrix metalloproteinase-13 (MMP-13) gene is induced by PTH in osteoblastic cells as a secondary response through the protein kinase A pathway requiring the runt domain and activator protein 1 binding sites of the proximal promoter. Here, we investigated the changes PTH causes in histone acetylation in this region (which contains the only deoxyribonuclease-hypersensitive sites in the promoter) leading to MMP-13 gene activation in these cells. Chromatin immunoprecipitation experiments revealed that PTH rapidly increased histone H4 acetylation followed by histone H3 acetylation associated with the different regions of the MMP-13 proximal promoter. The hormone also stimulated p300 histone acetyl transferase activity and increased p300 bound to the MMP-13 proximal promoter, and this required protein synthesis. Upon PTH treatment, Runx2, already bound to the runt domain site of the MMP-13 promoter, interacted with p300, which then acetylated histones H4 and H3. The knockdown of either Runx2 or p300 by RNA interference reduced PTH-induced acetylation of histones H3 and H4, association of p300 with the MMP-13 promoter, and resultant MMP-13 gene transcription. Overall, our studies suggest that without altering the gross chromatin structure, PTH stimulates acetylation of histones H3 and H4 via recruitment of p300 to Runx2 bound to the MMP-13 promoter, resulting in gene activation. This work establishes the molecular basis of transcriptional regulation in osteoblasts by PTH, a hormone acting through a G-protein coupled receptor.
Department of Physiology and Biophysics, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
Christine E
CE
Boumah
Minnkyong
M
Lee
Nagarajan
N
Selvamurugan
Emi
E
Shimizu
Nicola C
NC
Partridge
D000107:Acetylation; D000818:Animals; D016723:Bone Remodeling; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism*; D050881:E1A-Associated p300 Protein / Q000378:metabolism; D006657:Histones / Q000737:chemistry* / Q000378:metabolism; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism*; D010006:Osteoblasts / Q000378:metabolism; D010281:Parathyroid Hormone / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D051381:Rats; D015533:Transcriptional Activation
D050659:Core Binding Factor Alpha 1 Subunit; D006657:Histones; D010281:Parathyroid Hormone; C493717:Runx2 protein, rat; D050881:E1A-Associated p300 Protein; C494362:Ep300 protein, rat; D053509:Matrix Metalloproteinase 13