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. 2015:2015:416751.
doi: 10.1155/2015/416751. Epub 2015 Sep 20.

Recognition Code of ZNF191(243-368) and Its Interaction with DNA

Dongxin Zhao  1 , Zhongxian Huang  2
Affiliations

Recognition Code of ZNF191(243-368) and Its Interaction with DNA

Dongxin Zhao et al. Bioinorg Chem Appl. 2015.

Abstract

ZNF191(243-368) is the C-terminal region of ZNF191 which contains a putative DNA-binding domain of four Cys2His2 zinc finger motifs. In this study, an expression vector of a fusion protein of ZNF191(243-368) with glutathione-S-transferase (GST) was constructed and transformed into Escherichia coli BL21. The fusion protein GST-ZNF191(243-368) was expressed using this vector to investigate the protein-DNA binding reaction through an affinity selection strategy on the basis of the binding quality of the zinc finger domain. Results showed that ZNF191(243-368) can selectively bind with sequences and react with genes which contain an AGGG core. However, the recognition mechanism of Cys2His2 zinc finger proteins to DNA warrants further investigation.

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Figures

Figure 1
Figure 1
SDS-PAGE of GST-ZNF191(243-368) fusion protein. Lane 1: protein purified by G75, lane 2: protein marker, and lane 3: protein purified by affinity resin.
Figure 2
Figure 2
Schematic representation of DNA selecting of GST-ZNF191(243-368). Glutathione sepharose 4B is bound by the GST of fusion protein. Meanwhile, the zinc finger protein binds its recognition sequence in DNA and thus immobilizes the DNA on glutathione sepharose 4B.
Figure 3
Figure 3
Alignment of 69 selected oligonucleotides bound to ZNF191(243-368). There are 51 sequences (74%) containing AGGG motif or similar AGGA, AGGT, AGGC motif, and the content of GC in others is high.
Figure 4
Figure 4
Different UV spectrum of GST-ZNF191(243-368) and GST. Solid line is UV spectrum of ZNF191(243-368) and dash line is different UV spectrum of GST-ZNF191(243-368) and GST.
Figure 5
Figure 5
Different CD spectrum of GST-ZNF191(243-368) and GST. Solid line is CD spectrum of ZNF191(243-368) and dash line is different CD spectrum of GST-ZNF191(243-368) and GST.
Figure 6
Figure 6
Fluorescence spectra of EB-DNA1 titrated by ZNF191(243-368). C DNA = 0.5 μmol·L−1, C EB = 1 μmol·L−1, from up to down, and C protein/C DNA = 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, respectively.
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References

    1. Berg J. M., Shi Y. The galvanization of biology: a growing appreciation for the roles of zinc. Science. 1996;271(5252):1081–1085. doi: 10.1126/science.271.5252.1081. - DOI - PubMed
    1. Razin S. V., Borunova V. V., Maksimenko O. G., Kantidze O. L. Cys2his2 zinc finger protein family: classification, functions, and major members. Biochemistry. 2012;77(3):217–226. doi: 10.1134/s0006297912030017. - DOI - PubMed
    1. Klug A. The discovery of zinc fingers and their development for practical applications in gene regulation and genome manipulation. Quarterly Reviews of Biophysics. 2010;43(1):1–21. doi: 10.1017/s0033583510000089. - DOI - PubMed
    1. Greisman H. A., Pabo C. O. A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites. Science. 1997;275(5300):657–661. doi: 10.1126/science.275.5300.657. - DOI - PubMed
    1. Wolfe S. A., Nekludova L., Pabo C. O. DNA recognition by Cys2His2 zinc finger proteins. Annual Review of Biophysics and Biomolecular Structure. 2000;29:183–212. doi: 10.1146/annurev.biophys.29.1.183. - DOI - PubMed

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