Nov, 2005
Double-stranded RNA-dependent protein kinase is required for bone calcification in MC3T3-E1 cells in vitro
EXPERIMENTAL CELL RESEARCH
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- Volume
- 311
- Number
- 1
- First page
- 117
- Last page
- 125
- Language
- English
- Publishing type
- Research paper (scientific journal)
- DOI
- 10.1016/j.yexcr.2005.09.006
- Publisher
- ELSEVIER INC
In this study, we demonstrated that double-stranded RNA-dependent protein kinase (PKR) is required for the calcification of osteoblasts via the signal transducers and activators of transcription 1 alpha(STAT1 alpha) signaling in vitro. A dominant-negative mutant PKR cDNA, in which the amino acid lysine at 296 was replaced with arginine and which does not have catalytic activity, was transfected into mouse ostcoblastic MC3T3-E1 cells, thereby, we established cells that stably expressed the PKR mutant gene (PKR-K/R). Phosphorylation of PKR was not stimulated by polyinosic-polycytidylic acid in the mutant cells. The PKR-K/R mutant cells exhibited up-regulated cell growth and had low alkaline phosphatase (ALP) activity. The PKR-K/R mutant cells were not able to form bone nodules in vitro. In the PKR-K/R mutant cells, runt-related gene 2 (Runx2)-mediated transcription decreased compared with the levels in the control cells. The expression of STAT1 alpha protein increased and the protein was translocated to the nucleus in the PKR-K/R mutant cells. When the expression of STAT1 alpha a protein in PKR Mutant cells was suppressed using RNAi, the activity of Runx2-mediated transcription recovered to the control level. Our results indicate that PKR is a stimulator of Runx2 transcription and is a negative modulator of STAT1 alpha expression. Our findings also suggest that PKR plays important roles in the differentiation and calcification of osteoblasts by modulating STAT1 alpha and/or Runx2 expression. (c) 2005 Elsevier Inc. All rights reserved.
- Link information
- ID information
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- DOI : 10.1016/j.yexcr.2005.09.006
- ISSN : 0014-4827
- Pubmed ID : 16216244
- Web of Science ID : WOS:000233268100011