Polycystin-1 regulates skeletogenesis through stimulation of the osteoblast-specific transcription factor RUNX2-II

Zhousheng Xiao, Shiqin Zhang, Brenda S. Magenheimer, Junming Luo, Leigh Quarles

Research output: Contribution to journalArticle

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Abstract

Polycystin-1 (PC1) may play an important role in skeletogenesis through regulation of the bone-specific transcription factor Runx2-II. In the current study we found that PC1 co-localizes with the calcium channel polycystin-2 (PC2) in primary cilia of MC3T3-E1 osteoblasts. To establish the role of Runx2-II in mediating PC1 effects on bone, we crossed heterozygous Pkd1m1Bei and Runx2-II mice to create double heterozygous mice (Pkd1+/m1Bei/Runx2- II+/-) deficient in both PC1 and Runx2-II. Pkd1+/m1Bei/ Runx2-II+/- mice exhibited additive reductions in Runx2-II expression that was associated with impaired endochondral bone development, defective osteoblast-mediated bone formation, and osteopenia. In addition, we found that basal intracellular calcium levels were reduced in homozygous Pkd1 m1Bei osteoblasts. In contrast, overexpression of a PC1 C-tail construct increased intracellular calcium and selectively stimulated Runx2-II P1 promoter activity in osteoblasts through a calcium-dependent mechanism. Site-directed mutagenesis of critical amino acids in the coiled-coil domain of PC1 required for coupling to PC2 abolished PC1-mediated Runx2-II P1 promoter activity. Additional promoter analysis mapped the PC1-responsive region to the "osteoblast-specific" enhancer element between -420 and -350 bp that contains NFI and AP-1 binding sites. Chromatin immunoprecipitation assays confirmed the calcium-dependent binding of NFI to this region. These findings indicate that PC1 regulates osteoblast function through intracellular calcium-dependent control of Runx2-II expression. The overall function of the primary cilium-polycystin complex may be to sense and transduce environmental clues into signals regulating osteoblast differentiation and bone development.

Original languageEnglish (US)
Pages (from-to)12624-12634
Number of pages11
JournalJournal of Biological Chemistry
Volume283
Issue number18
DOIs
StatePublished - May 2 2008

Fingerprint

Osteoblasts
Transcription Factors
Bone
Calcium
Cilia
Bone Development
TRPP Cation Channels
polycystic kidney disease 1 protein
Bone and Bones
Mutagenesis
Metabolic Bone Diseases
Chromatin Immunoprecipitation
Transcription Factor AP-1
Calcium Channels
Site-Directed Mutagenesis
Osteogenesis
Chromatin
Tail
Assays
Binding Sites

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Polycystin-1 regulates skeletogenesis through stimulation of the osteoblast-specific transcription factor RUNX2-II. / Xiao, Zhousheng; Zhang, Shiqin; Magenheimer, Brenda S.; Luo, Junming; Quarles, Leigh.

In: Journal of Biological Chemistry, Vol. 283, No. 18, 02.05.2008, p. 12624-12634.

Research output: Contribution to journalArticle

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abstract = "Polycystin-1 (PC1) may play an important role in skeletogenesis through regulation of the bone-specific transcription factor Runx2-II. In the current study we found that PC1 co-localizes with the calcium channel polycystin-2 (PC2) in primary cilia of MC3T3-E1 osteoblasts. To establish the role of Runx2-II in mediating PC1 effects on bone, we crossed heterozygous Pkd1m1Bei and Runx2-II mice to create double heterozygous mice (Pkd1+/m1Bei/Runx2- II+/-) deficient in both PC1 and Runx2-II. Pkd1+/m1Bei/ Runx2-II+/- mice exhibited additive reductions in Runx2-II expression that was associated with impaired endochondral bone development, defective osteoblast-mediated bone formation, and osteopenia. In addition, we found that basal intracellular calcium levels were reduced in homozygous Pkd1 m1Bei osteoblasts. In contrast, overexpression of a PC1 C-tail construct increased intracellular calcium and selectively stimulated Runx2-II P1 promoter activity in osteoblasts through a calcium-dependent mechanism. Site-directed mutagenesis of critical amino acids in the coiled-coil domain of PC1 required for coupling to PC2 abolished PC1-mediated Runx2-II P1 promoter activity. Additional promoter analysis mapped the PC1-responsive region to the {"}osteoblast-specific{"} enhancer element between -420 and -350 bp that contains NFI and AP-1 binding sites. Chromatin immunoprecipitation assays confirmed the calcium-dependent binding of NFI to this region. These findings indicate that PC1 regulates osteoblast function through intracellular calcium-dependent control of Runx2-II expression. The overall function of the primary cilium-polycystin complex may be to sense and transduce environmental clues into signals regulating osteoblast differentiation and bone development.",
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