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

Dongxin Zhao,Zhongxian Huang

doi:10.1155/2015/416751

Dongxin Zhao, Zhongxian Huang

Open Access

https://doi.org/10.1155/2015/416751

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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.

Highlights

Kruppel-type (C2H2) zinc finger is ubiquitous motif which mediates the sequence-specific recognition of DNA and widely exists in eukaryotes
The present study aims to elucidate the function of ZNF191(243-368) at the protein level
After the purified fusion protein was bound to the resin, the mixture was used for the following binding experiment

Summary

Introduction

Kruppel-type (C2H2) zinc finger is ubiquitous motif which mediates the sequence-specific recognition of DNA and widely exists in eukaryotes This zinc finger protein can bind DNA, RNA, or other proteins and assume critical roles in various biological functions, including cell differentiation and embryo development [1–3]. Previous studies explored the crystal/NMR structures and DNA-binding sites of zinc finger proteins, and some works reported the possibility of previewing the recognition site of a novel C2H2 zinc finger protein by sequence analysis [4–7]. These observations are insufficient to affirm the recognition code of each amino acid residue [8, 9]. Results indicated that the consensus binding site of ZNF191(243-368) had an “AGGG” core and implicated that the chemical rules for the sequence-specific recognition of Cys2His to DNA should be used cautiously

Materials and Methods

Results and Discussion

Conclusion