Developmental profiles of phosphorylated and unphosphorylated CREBs in murine calvarial MC3T3-E1 cells

Journal of Biochemistry
M K SakamotoS Nakamura

Abstract

The cAMP-responsive element (CRE) binding protein/activating transcription factor (CREB/ATF) family plays a major role in the expression of skeletal-specific genes and skeletal tissue development. We analyzed the changes of the amount, degree of phosphorylation and binding activity of the CREB/ATF family in the course of development of the murine calvarial osteoblastic cell line MC3T3-E1 as an in vitro model system of bone formation. The amount of CREB in the whole-cell extract detectable by Western blot analysis was high through all stages of development and maximal in the proliferation stage. The degree of phosphorylation estimated with anti-phosphorylated CREB antibody changed greatly and reached high levels in the proliferation stage and early mineralization stage. The ratio of phosphorylated CREB to total CREB in the CREB-CRE complex was also examined by gel shift assay. Although the binding to the consensus/CRE probe reached almost equally high levels in the proliferation stage and early mineralization stage, the relative level of phosphorylated CREB in the CREB-CRE complex was different in these two stages. In the early mineralization stage, most CREB bound to consensus/CRE was phosphorylated, while both phosphorylated a...Continue Reading

Citations

Jul 22, 2009·Journal of Bone and Mineral Research : the Official Journal of the American Society for Bone and Mineral Research·Laura M YergesUNKNOWN Osteoporotic Fractures in Men (MrOS) Study Group
Dec 24, 2005·The Journal of Biological Chemistry·Noritaka MatsuoFrancesco Ramirez
Dec 3, 1999·Journal of Neurochemistry·P YuanH K Manji

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