- Research Article
Identification of the DNA binding element of the human ZNF300 protein
Cellular & Molecular Biology Letters volume 13, pages 391–403 (2008)
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.
electrophoretic mobility shift assay
IL-2 receptor beta chain
zinc finger 300
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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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
- Zinc finger
- DNA binding