Skip to main content
Download PDF
  • Research Article
  • Published:

Identification of the DNA binding element of the human ZNF300 protein

Cellular & Molecular Biology Letters volume 13, pages 391–403 (2008)Cite this article

Abstract

The human ZNF300 gene is a member of the KRAB/C2H2 zinc finger gene family, the members of which are known to be involved in various developmental and pathological processes. Here, we show that the ZNF300 gene encodes a 68-kDa nuclear protein that binds DNA in a sequence-specific manner. The ZNF300 DNA binding site, C(t/a)GGGGG(c/g)G, was defined via a random oligonucleotide selection assay, and the DNA binding site was further confirmed by electrophoretic mobility shift assays. A potential ZNF300 binding site was found in the promoter region of the human IL-2Rβ gene. The results of electrophoretic mobility shift assays indicated that ZNF300 bound to the ZNF300 binding site in the IL-2Rβ promoter in vitro. Transient co-transfection assays showed that ZNF300 could activate the IL-2Rβ promoter, and that the activation was abrogated by the mutation of residues in the ZNF300 binding site. Identifying the DNA binding site and characterizing the transcriptional regulation property of ZNF300 would provide critical insights into its potential as a transcriptional regulator.

Abbreviations

EMSA:

electrophoretic mobility shift assay

IL-2Rβ:

IL-2 receptor beta chain

KRAB:

Krüppel-associated box

TK:

thymidine kinase

ZNF300:

zinc finger 300

References

  1. Schuh, R., Aicher, W., Gaul, U., Cote, S., Preiss, A., Maier, D., Seifert, E., Nauber, U., Schroder, C. and Kemler, R. A conserved family of nuclear proteins containing structural elements of the finger protein encoded by Kruppel, a Drosophila segmentation gene. Cell 47 (1986) 1025–1032.

    Article  PubMed  CAS  Google Scholar 

  2. Dai, K.S. and Liew, C.C. Chromosomal, in silico and in vitro expression analysis of cardiovascular-based genes encoding zinc finger proteins. J. Mol. Cell. Cardiol. 31 (1999) 1749–1769.

    Article  PubMed  CAS  Google Scholar 

  3. Decker, E.L., Nehmann, N., Kampen, E., Eibel, H., Zipfel, P.F. and Skerka, C. Early growth response proteins (EGR) and nuclear factors of activated T cells (NFAT) form heterodimers and regulate proinflammatory cytokine gene expression. Nucleic Acids Res. 31 (2003) 911–921.

    Article  PubMed  CAS  Google Scholar 

  4. Gou, D.M., Wang, J., Gao, L., Sun, Y., Peng, X., Huang, J. and Li, W.X. Identification and functional analysis of a novel human KRAB/C2H2 zinc finger gene ZNF300. Biochim. Biophys. Acta 1676 (2004) 203–209.

    PubMed  CAS  Google Scholar 

  5. Sharon, M., Gnarra, J.R. and Leonard, W.J. A 100-kilodalton protein is associated with the murine interleukin 2 receptor: biochemical evidence that p100 is distinct from the alpha and beta chains. Proc. Natl. Acad. Sci. USA 87 (1990) 4869–4873.

    Article  PubMed  CAS  Google Scholar 

  6. Suzuki, H., Duncan, G.S., Takimoto, H. and Mak, T.W. Abnormal development of intestinal intraepithelial lymphocytes and peripheral natural killer cells in mice lacking the IL-2 receptor beta chain. J. Exp. Med. 185 (1997) 499–505.

    Article  PubMed  CAS  Google Scholar 

  7. Suzuki, H., Kundig, T.M., Furlonger, C., Wakeham, A., Timms, E., Matsuyama, T., Schmits, R., Simard, J.J., Ohashi, P.S., Griesser, H. Deregulated T-cell activation and autoimmunity in mice lacking interleukin-2 receptor beta. Science 268 (1995) 1472–1476.

    Article  PubMed  CAS  Google Scholar 

  8. Espinoza-Delgado, I., Ortaldo, J.R., Winkler-Pickett, R., Sugamura, K., Varesio, L. and Longo, D.L. Expression and role of p75 interleukin 2 receptor on human monocytes. J. Exp. Med. 171 (1990) 1821–1826.

    Article  PubMed  CAS  Google Scholar 

  9. Pierrou, S., Enerback, S. and Carlsson, P. Selection of high-affinity binding sites for sequence-specific, DNA binding proteins from random sequence oligonucleotides. Anal. Biochem. 229 (1995) 99–105.

    Article  PubMed  CAS  Google Scholar 

  10. Zweidler-Mckay, P.A., Grimes, H.L., Flubacher, M.M. and Tsichlis, P.N. Gfi-1 encodes a nuclear zinc finger protein that binds DNA and functions as a transcriptional repressor. Mol. Cell. Biol. 16 (1996) 4024–4034.

    PubMed  CAS  Google Scholar 

  11. Kim, J.C., Yoon, J.B., Koo, H.S. and Chung, I.K. Cloning and characterization of the 5′-flanking region for the human topoisomerase III gene. J. Biol. Chem. 273 (1998) 26130–26137.

    Article  PubMed  CAS  Google Scholar 

  12. Wolfe, S.A., Nekludova, L. and Pabo, C.O. DNA recognition by Cys2His2 zinc finger proteins. Annu. Rev. Biophys. Biomol. Struct. 29 (2000) 183–212.

    Article  PubMed  CAS  Google Scholar 

  13. Mendez-Vidal, C., Wilhelm, M.T., Hellborg, F., Qian, W. and Wiman, K.G. The p53-induced mouse zinc finger protein wig-1 binds double-stranded RNA with high affinity. Nucleic Acids Res. 30 (2002) 1991–1996.

    Article  PubMed  CAS  Google Scholar 

  14. Finerty, P.J., Jr. and Bass, B.L. A Xenopus zinc finger protein that specifically binds dsRNA and RNA-DNA hybrids. J. Mol. Biol. 271 (1997) 195–208.

    Article  PubMed  CAS  Google Scholar 

  15. O’Neill, E.A., Fletcher, C., Burrow, C.R., Heintz, N., Roeder, R.G. and Kelly, T.J. Transcription factor OTF-1 is functionally identical to the DNA replication factor NF-III. Science 241 (1988) 1210–1213.

    Article  PubMed  CAS  Google Scholar 

  16. Lin, J.X., Bhat, N.K., John, S., Queale, W.S. and Leonard, W.J. Characterization of the human interleukin-2 receptor beta-chain gene promoter: regulation of promoter activity by ets gene products. Mol. Cell. Biol. 13 (1993) 6201–6210.

    PubMed  CAS  Google Scholar 

  17. Wang, S., Liu, S., Wu, M.H., Geng, Y. and Wood, C. Identification of a cellular protein that interacts and synergizes with the RTA (ORF50) protein of Kaposi’s sarcoma-associated herpesvirus in transcriptional activation. J. Virol. 75 (2001) 11961–11973.

    Article  PubMed  CAS  Google Scholar 

  18. Qi, X., Li, Y., Xiao, J., Yuan, W., Yan, Y., Wang, Y., Liang, S., Zhu, C., Chen, Y., Liu, M. and Wu, X. Activation of transcriptional activities of AP-1 and SRE by a new zinc-finger protein ZNF641. Biochem. Biophys. Res. Commun. 339 (2006) 1155–1164.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, 430072, P.R. China

    Hongling Qiu, Lu Xue, Li Gao, Huanjie Shao, Di Wang, Mingxiong Guo & Wenxin Li

Authors
  1. Hongling Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lu Xue
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Li Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huanjie Shao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Di Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mingxiong Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wenxin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wenxin Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Qiu, H., Xue, L., Gao, L. et al. Identification of the DNA binding element of the human ZNF300 protein. Cell Mol Biol Lett 13, 391–403 (2008). https://doi.org/10.2478/s11658-008-0005-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-008-0005-x

Key words

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us