Skip to main content
SpringerLink
Log in

Construction and functional analysis of novel dominant-negative mutant of human SOX18 protein

  • Published:
Biochemistry (Moscow) Aims and scope Submit manuscript

Abstract

SOX18 transcription factor plays important roles in a range of biological processes such as vasculogenesis, hair follicle development, lymphangiogenesis, atherosclerosis, and angiogenesis. In this paper we present the generation of a novel SOX18 dominant-negative mutant (SOX18DN) encoding truncated SOX18 protein that lacks a trans-activation domain. We show that both wild-type SOX18 (SOX18wt) and truncated human SOX18 proteins are able to bind to their consensus sequence in vitro. Functional analysis confirmed that SOX18wt has potent trans-activation properties, while SOX18DN displays dominant-negative effect. We believe that these SOX18wt and SOX18DN expression constructs could be successfully used for further characterization of the function of this protein.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

HMG:

high mobility group box

SOX18wt :

wild-type SOX18

SOX18DN:

truncated dominant-negative mutant protein SOX18

Sry gene:

sex determining region of Y chromosome

TAD:

trans-activation domain (of SOX18)

References

  1. Pevny, L. H., and Lovell-Badge, R. (1997) Curr. Opin. Genet. Dev., 7, 338–344.

    Article  PubMed  CAS  Google Scholar 

  2. Wegner, M. (1999) Nucleic Acids Res., 27, 1409–1420.

    Article  PubMed  CAS  Google Scholar 

  3. Bowles, J., Schepers, G., and Koopman, P. (2000) Dev. Biol., 227, 239–255.

    Article  PubMed  CAS  Google Scholar 

  4. Hosking, B. M., Muscat, G. E., Koopman, P. A., Dowhan, D. H., and Dunn, T. L. (1995) Nucleic Acids Res., 23, 2626–2628.

    Article  PubMed  CAS  Google Scholar 

  5. Sandholzer, J., Hoeth, M., Piskacek, M., Mayer, H., and de Martin, R. (2007) Biochem. Biophys. Res. Commun., 360, 370–374.

    Article  PubMed  CAS  Google Scholar 

  6. Cermenati, S., Moleri, S., Cimbro, S., Corti, P., Del Giacco, L., Amodeo, R., Dejana, E., Koopman, P., Cotelli, F., and Beltrame, M. (2008) Blood, 111, 2657–2666.

    Article  PubMed  CAS  Google Scholar 

  7. Matsui, T., Kanai-Azuma, M., Hara, K., Matoba, S., Hiramatsu, R., Kawakami, H., Kurohmaru, M., Koopman, P., and Kanai, Y. (2006) J. Cell Sci., 119, 3513–3526.

    Article  PubMed  CAS  Google Scholar 

  8. Fawcett, S. R., and Klymkowsky, M. W. (2004) Gene Expr. Patterns, 4, 29–33.

    Article  PubMed  CAS  Google Scholar 

  9. Olsson, J. E., Kamachi, Y., Penning, S., Muscat, G. E., Kondoh, H., and Koopman, P. (2001) Gene, 271, 151–158.

    Article  PubMed  CAS  Google Scholar 

  10. James, K., Hosking, B., Gardner, J., Muscat, G. E., and Koopman, P. (2003) Genesis, 36, 1–6.

    Article  PubMed  CAS  Google Scholar 

  11. Pennisi, D., Gardner, J., Chambers, D., Hosking, B., Peters, J., Muscat, G., Abbott, C., and Koopman, P. (2000) Nat. Genet., 24, 434–437.

    Article  PubMed  CAS  Google Scholar 

  12. Irrthum, A., Devriendt, K., Chitayat, D., Matthijs, G., Glade, C., Steijlen, P. M., Fryns, J. P., Van Steensel, M. A., and Vikkula, M. (2003) Am. J. Hum. Genet., 72, 1470–1478.

    Article  PubMed  CAS  Google Scholar 

  13. Pennisi, D., Bowles, J., Nagy, A., Muscat, G., and Koopman, P. (2000) Mol. Cell Biol., 20, 9331–9336.

    Article  PubMed  CAS  Google Scholar 

  14. Herpers, R., van de Kamp, E., Duckers, H. J., and Schulte-Merker, S. (2008) Circ. Res., 102, 12–15.

    Article  PubMed  CAS  Google Scholar 

  15. Miyagi, S., Kato, H., and Okuda, A. (2009) Cell. Mol. Life Sci., 66, 3675–3684.

    Article  PubMed  CAS  Google Scholar 

  16. Smits, P., Li, P., Mandel, J., Zhang, Z., Deng, J. M., Behringer, R. R., de Crombrugghe, B., and Lefebvre, V. (2001) Dev. Cell., 1, 277–290.

    Article  PubMed  CAS  Google Scholar 

  17. Barrionuevo, F., and Scherer, G. (2010) Int. J. Biochem. Cell Biol., 42, 433–436.

    Article  PubMed  CAS  Google Scholar 

  18. Chaboissier, M. C., Kobayashi, A., Vidal, V. I., Lutzkendorf, S., van de Kant, H. J., Wegner, M., de Rooij, D. G., Behringer, R. R., and Schedl, A. (2004) Development, 131, 1891–1901.

    Article  PubMed  CAS  Google Scholar 

  19. Stanojcic, S., and Stevanovic, M. (2000) Biochim. Biophys. Acta, 1492, 237–241.

    Article  PubMed  CAS  Google Scholar 

  20. Kuhlbrodt, K., Herbarth, B., Sock, E., Hermans-Borgmeyer, I., and Wegner, M. (1998) J. Neurosci., 18, 237–250.

    PubMed  CAS  Google Scholar 

  21. Dignam, J. D., Lebovitz, R. M., and Roeder, R. G. (1983) Nucleic Acids Res., 11, 1475–1489.

    Article  PubMed  CAS  Google Scholar 

  22. Kovacevic Grujicic, N., Mojsin, M., Krstic, A., and Stevanovic, M. (2005) Gene, 344, 287–297.

    Article  PubMed  CAS  Google Scholar 

  23. Milivojevic, M., Nikcevic, G., Kovacevic-Grujicic, N., Krstic, A., Mojsin, M., Drakulic, D., and Stevanovic, M. (2010) Arch. Biol. Sci., Belgrade, 62, 199–210.

    Article  Google Scholar 

  24. Li, J., Pan, G., Cui, K., Liu, Y., Xu, S., and Pei, D. (2007) J. Biol. Chem., 282, 19481–19492.

    Article  PubMed  CAS  Google Scholar 

  25. Shih, Y. H., Kuo, C. L., Hirst, C. S., Dee, C. T., Liu, Y. R., Laghari, Z. A., and Scotting, P. J. (2010) Development, 137, 2671–2681.

    Article  PubMed  CAS  Google Scholar 

  26. Cossais, F., Wahlbuhl, M., Kriesch, J., and Wegner, M. (2010) Hum. Mol. Genet., 19, 2409–2420.

    Article  PubMed  CAS  Google Scholar 

  27. Fontijn, R. D., Volger, O. L., Fledderus, J. O., Reijerkerk, A., de Vries, H. E., and Horrevoets, A. J. (2008) Am. J. Physiol. Heart Circ. Physiol., 294, H891–900.

    Article  PubMed  CAS  Google Scholar 

  28. Kelberman, D., Rizzoti, K., Avilion, A., Bitner-Glindzicz, M., Cianfarani, S., Collins, J., Chong, W. K., Kirk, J. M., Achermann, J. C., Ross, R., Carmignac, D., Lovell-Badge, R., Robinson, I. C., and Dattani, M. T. (2006) J. Clin. Invest., 116, 2442–2455.

    PubMed  CAS  Google Scholar 

  29. Chen, Y., Shi, L., Zhang, L., Li, R., Liang, J., Yu, W., Sun, L., Yang, X., Wang, Y., Zhang, Y., and Shang, Y. (2008) J. Biol. Chem., 283, 17969–17978.

    Article  PubMed  CAS  Google Scholar 

  30. Francois, M., Caprini, A., Hosking, B., Orsenigo, F., Wilhelm, D., Browne, C., Paavonen, K., Karnezis, T., Shayan, R., Downes, M., Davidson, T., Tutt, D., Cheah, K. S., Stacker, S. A., Muscat, G. E., Achen, M. G., Dejana, E., and Koopman, P. (2008) Nature, 456, 643–647.

    Article  PubMed  CAS  Google Scholar 

  31. Garcia-Ramirez, M., Martinez-Gonzalez, J., Juan-Babot, J. O., Rodriguez, C., and Badimon, L. (2005) Arterioscler. Thromb. Vasc. Biol., 25, 2398–2403.

    Article  PubMed  CAS  Google Scholar 

  32. Young, N., Hahn, C. N., Poh, A., Dong, C., Wilhelm, D., Olsson, J., Muscat, G. E., Parsons, P., Gamble, J. R., and Koopman, P. (2006) J. Natl. Cancer Inst., 98, 1060–1067.

    Article  PubMed  CAS  Google Scholar 

  33. Luo, M., Guo, X. T., Yang, W., Liu, L. Q., Li, L. W., and Xin, X. Y. (2008) Med. Hypotheses, 70, 880–882.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, PO BOX 23, 11010, Belgrade, Serbia

    M. Milivojevic, I. Petrovic, N. Kovacevic-Grujicic, J. Popovic, M. Mojsin & M. Stevanovic

Authors
  1. M. Milivojevic
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I. Petrovic
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N. Kovacevic-Grujicic
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Popovic
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Mojsin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. Stevanovic
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to I. Petrovic.

Additional information

Published in Russian in Biokhimiya, 2013, Vol. 78, No. 11, pp. 1635–1642.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Milivojevic, M., Petrovic, I., Kovacevic-Grujicic, N. et al. Construction and functional analysis of novel dominant-negative mutant of human SOX18 protein. Biochemistry Moscow 78, 1287–1292 (2013). https://doi.org/10.1134/S0006297913110096

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0006297913110096

Key words

Search

Navigation