International Journal of Oral Biology

  • 제32권4호
  • /
  • Pages.119-125
  • /
  • 2007
  • /
  • 1226-7155(pISSN)
  • /
  • 2287-6618(eISSN)
대한구강생물학회 (The Korean Academy of Oral Biology)
권호 표지

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)
  • 발행 : 2007.12.31
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

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.

키워드

참고문헌

  1. Benson, M. D., Bargeon, J., Xiao, G., Thomas, P. E., Kim, A., Cui, Y. and Franceschi, R. T.: Identification of a homeodomain binding element in the bone sialoprotein gene promoter that is required for its osteoblastic gene promoter that is required for its osteoblastic-selective expression. J. Biol. Chem. 275: 13907-13917, 2000 https://doi.org/10.1074/jbc.275.18.13907
  2. Bryan, J. T. and Morasso, M. I.: The Dlx3 protein harbors basic residues required for nuclear localization. transcriptional activity and binding to Msxl. J. Cell Sci. 113:4013-4023, 2000
  3. Cha, J. H., Ryoo, H. M., Woo, K. M., Kim, G. S. and Baek, J. H.: Dlx3 plays a role as a positive regulator of osteoclast differentiation. Int. J. Oral Biol. 32:85-91, 2007
  4. Choi, S. J., Song, I. S., Ryu, O. H., Choi, S. W., Hart, P. S., Wu, W. W., Shen, R. F. and Hart, T. C; A4 bp deletion mutation in DLX3 enhances osteoblastic differentiation and bone formation in vitro. Bone (2007), doi;10.1016/j. bone. 2007. 08.047
  5. Feledy, J. A., Morasso, M. I., Jang, S. I. and Sargent, T. D.: Transcriptional activation by the homeodomain protein distal-less 3. Nucleic Acids Res. 27:764-770, 1999 https://doi.org/10.1093/nar/27.3.764
  6. Ghanem, N., Jarinova, O., Amores, A., Long, Q., Hatch, G., Park, B. K., Rubenstein, J. L. R. and Ekker, M.: Regulatory roles of conserved intergenic domains in vertebrate Dlx bigene clusters. Genome Res. 13:533-543, 2003 https://doi.org/10.1101/gr.716103
  7. Hassan, M. Q., Javed, A, Morasso, M. I., Karlin, J., Montecino, M., Wijnen, A. J. V.. Stein, G. S., Stein, J. L. and Lian, J. B.: Dlx3 transcriptional regulation of osteoblast differentiation: temporal recruitment of Msx2, Dlx3 and DlxS homeodomain proteins to chromatin of the osteocalcin gene. Mol. Cell. Biol. 24:9248-9261, 2004 https://doi.org/10.1128/MCB.24.20.9248-9261.2004
  8. Hassan, M. Q., Tare, R. S., Lee, S. H., Mandeville, M., Morasso, M. I., Javed, A., van Wijnen, A. J., Stein, J. L., Stein, G. S. and Lian, J. B.: BMP2 commitment to the osteogenic lineage involves activation of Runx2 by DLX3 and a homeodomain transcriptional network. J. Biol. Chem. 281:40515-40526, 2006 https://doi.org/10.1074/jbc.M604508200
  9. Kim, Y. J., Lee, M. H., Wozney, J. M., Cho, J. Y. and Ryoo, H. M.: Bone morphogenetic protein- 2- induced alkaline phosphatase expression is stimulated by Dlx5 and repressed by Msx2. J. Biol. Chem. 279:50773-50780, 2004 https://doi.org/10.1074/jbc.M404145200
  10. Lacey, D. L., Timms, E., Tan, H. L., Kelley, M. J., Dunstan, C. R., Burgess, T., Elliott, R., Colombero, A., Elliot, G., Scully, S., Hsu, H., Sullivan, J., Hawkins, N., Davy, E., Capparelli, C., Eli, A., Qian, Y. X., Kaufman, S., Sarosi, I., Shalhoub, V.. Senaldi, G., Guo, J., Delaney, J. and Boyle, W.J.: Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93: 165-176, 1998 https://doi.org/10.1016/S0092-8674(00)81569-X
  11. Laughon, A: DNA binding specificity of homeodomains. Biochemistry 30: 11357 -11367, 1991 https://doi.org/10.1021/bi00112a001
  12. McGinnis, W. and Krumlauf, R.: Homeobox genes and axial patterning. Cell 68:283-302, 1992 https://doi.org/10.1016/0092-8674(92)90471-N
  13. Panganiban, G. and Rubenstein, J. L.: Developmental functions of the Distal-less/Dlx homeobox genes. Development 129:4371-4386, 2002
  14. Park, G. T. and Morasso, M. I.: Regulation of the Dlx3 homeobox gene upon differentiation of mouse keratinocytes. J. Biol. Chem. 274:26599-26608, 1999 https://doi.org/10.1074/jbc.274.37.26599
  15. Price, J. A., Bowden, D. W., Wright, J. T., Pettenati, M. J. and Hart, T. C: Identification of a mutation in DLX3 associated with tricho-dento-osseous (TDO) syndrome. Hum. Mol. Genet. 7:563-569, 1998a https://doi.org/10.1093/hmg/7.3.563
  16. Price, J. A., Wright, J. T., Kula, K., Bowden, D. W. and Hart, T. C.: A common DLX3 gene mutation is responsible for trichodento-osseous syndrome in Virginia and North Carolina families. J. Med. Genet. 35:825-828, 1998b https://doi.org/10.1136/jmg.35.10.825
  17. Rechsteiner, M. and Rogers, S. W.: PEST sequences and regulation by proteolysis. Trends Biochem. Sci. 21:267-271, 1996 https://doi.org/10.1016/S0968-0004(96)10031-1
  18. Rogers, S., Wells, R. and Rechsteiner, M.: Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science 234:364-368, 1986 https://doi.org/10.1126/science.2876518
  19. Scott, M. P., Tamkun, J. W. and Hartzell, G. W.: The structure and function of the homeodomain. Biochim. Biophys. Acta. 989:25-48, 1989
  20. Wright, J. T., Kula, K., Hall, K., Simmons, J. H. and Hart, T. C: Analysis of the tricho-dento-osseous syndrome genotype and phenotype. Am. J. Med. Genet. 72:197-204, 1997 https://doi.org/10.1002/(SICI)1096-8628(19971017)72:2<197::AID-AJMG14>3.0.CO;2-I
  21. Zhang, H., Hu, G., Wang, H., Sciavolino, P., Iler, N., Shen, M. M. and Abate-Shen, C.: Heterodimerization of Msx and Dlx homeoproteins results in functional antagonism. Mol. Cell. Biol. 17:2920-2932, 1997 https://doi.org/10.1128/MCB.17.5.2920