Focal Adhesion Kinase Activity Is Required for Bone Morphogenetic Protein—Smad1 Signaling and Osteoblastic Differentiation in Murine MC3T3-E1 Cells

• DOI: 10.1359/jbmr.2001.16.10.1772
• Publication Date: 2006-04-27T03:14:25Z
• Authors: Yasuhiro Tamura; Yasuhiro Takeuchi; Miyuki Suzawa; Seiji Fukumoto; Mitsuyasu Kato; Kohei Miyazono; Toshiro Fujita
• Keywords: 0301 basic medicine; Focal Adhesions; Osteoblasts; Bone Morphogenetic Protein 2; Cell Differentiation; Smad Proteins; Protein-Tyrosine Kinases; Phosphoproteins; Actins; Cell Line; Smad1 Protein; DNA-Binding Proteins; Actin Cytoskeleton; Mice; 03 medical and health sciences; Focal Adhesion Kinase 1; Focal Adhesion Protein-Tyrosine Kinases; Bone Morphogenetic Proteins; Animals; Humans; Signal Transduction; Smad4 Protein

Abstract Cell-matrix interactions via integrins are essential for osteoblastic differentiation. We have shown that signals activated by aggregation of α2β1-integrin with type I collagen are involved in the differentiation of osteoblastic MC3T3-E1 cells. Focal adhesion kinase (FAK) is an immediate downstream signal of the β1-integrin, and inactivation of FAK has been shown to disrupt osteoblastic differentiation. To elucidate roles of FAK in osteoblastic cells, we examined MC3T3-E1 cells stably expressing antisense FAK (asFAK) messenger RNA (mRNA). Alkaline phosphatase (ALP) activity, an osteoblastic marker, did not increase in asFAK cells with a long-term culture until 21 days or in response to bone morphogenetic protein 2 (BMP-2). Treatment with BMP-2 also failed to stimulate the expression of osteocalcin in asFAK cells. In control MC3T3-E1 cells, BMP-2 induced translocation of Smad1 into nuclei to stimulate transcriptional activity of the Smad6 promoter gene that contains a Smad1 response element. In contrast, BMP-2 failed to increase transcriptional activity of Smad6 promoter gene in asFAK cells, although it induced nuclear translocation of Smad1. These results indicate that FAK was involved in Smad1-dependent transcriptional activity but not in nuclear translocation of Smad1 in osteoblastic cells. Hence, FAK activation by integrins might converge transcriptional activation by BMP of its target genes in osteoblastic cells. These observations suggest that FAK activity is essential for BMP-Smad signaling to stimulate osteoblastic differentiation.