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Tricho-dento-osseous Syndrome Mutant Dlx3 Shows Lower Transactivation Potential but Has Longer Half-life than Wild-type Dlx3  

Cha, Ji-Hun (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University)
Ryoo, Hyun-Mo (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University)
Woo, Kyung-Mi (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University)
Kim, Gwan-Shik (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University)
Baek, Jeong-Hwa (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University)
Publication Information
International Journal of Oral Biology / v.32, no.4, 2007 , pp. 119-125 More about this Journal
Abstract
Dlx3 is a homeodomain protein and is known to play a role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM #190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. The molecular mechanisms that explain the phenotypic characteristics of TDO syndrome have not been clearly determined. In this study, we examined phenotypic characteristics of wild type DLX3(wtDlx3) and 4-BP DEL DLX3 (TDO mtDlx3) in C2C12 cells. To investigate how wtDlx3 and TDO mtDlx3 differentially regulate osteoblastic differentiation, reporter assays were performed by using luciferase reporters containing the promoters of alkaline phosphatase, bone sialoprotein or osteocalcin. Both wtDlx3 and TDO mtDlx3 enhanced significantly all the reporter activities but the effect of mtDlx3 was much weaker than that of wtDlx3. In spite of these differences in reporter activity, electrophoretic mobility shift assay showed that both wtDlx3 and TDO mtDlx3 formed similar amounts of DNA binding complexes with Dlx3 binding consensus sequence or with ALP promoter oligonucleotide bearing the Dlx3 binding core sequence. TDO mtDlx3 exhibits a longer half-life than wtDlx3 and it corresponds to PESTfind analysis result showing that potential PEST sequence was missed in carboxy terminal of TDO mtDlx3. In addition, co-immunoprecipitation demonstrated that TDO mtDlx3 binds to Msx2 more strongly than wtDlx3. Taken together, though TDO mtDlx3 acted as a weaker transcriptional activator than wtDlx3 in osteoblastic cells, there is possibility that during in vivo osteoblast differentiation TDO mtDlx3 may antagonize transcriptional repressor activity of Msx2 more effectively and for longer period than wtDlx3, resulting in enhancement of osteoblast differentiation.
Keywords
Dlx3; TDO mutant Dlx3; Osteoblast differentiation; Transactivation potential; Protein stabilization; Msx2;
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