Kaiso uses all three zinc fingers and adjacent sequence motifs for high affinity binding to sequence-specific and methyl-CpG DNA targets

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• 作者：Bethany A. Buck-Koehntop¹ ; Maria A. Martinez-Yamout¹ ; H. Jane Dyson ; Peter E. Wright ; ^{wright@scripps.edu}
• 关键词：MBP ; methyl-CpG DNA binding protein ; KBS ; Kaiso binding site ; ZF ; zinc finger ; NMR ; nuclear magnetic resonance ; HSQC ; heteronuclear single-quantum coherence ; EMSA ; electrophoretic gel mobility shift assays
• 刊名：FEBS Letters
• 出版年：2012
• 期刊代码：70_00145793
• 类别：bio
• 出版时间：23 March, 2012
• 卷：586
• 期：6
• 页码：734-739
• 文件大小：1223 K

摘要

Kaiso is a Cys₂His₂ zinc finger (ZF) protein that mediates methyl-CpG-dependent and sequence-specific transcriptional repression. As a first step towards elucidating the structural and molecular basis for recognition of these disparate DNA sequences, the minimal binding region of Kaiso was identified and optimal DNA sequences for high-affinity interactions were characterized. Contrary to previous findings, Kaiso requires all three zinc fingers plus adjacent protein regions for DNA recognition. An N-terminal extension contributes to structural stability, while an extended C-terminal region augments DNA binding. Complexes formed between the optimized Kaiso construct and both DNA sequences are suitable for future structural evaluation.