The Cys2His2 zinc finger domain is a well-defined compact structure stabilized by a coordinated zinc atom. Many proteins utilize tandem arrays of these concerted motifs to elicit specific recognition of nucleic acid sequences. While structurally conserved, this general fold can be utilized to distinguish diverse biological targets, requiring a deeper understanding for the molecular basis of nucleic acid recognition exhibited by these motifs. Our laboratory investigates a family of Cys2His2 zinc finger methyl-CpG binding proteins (MBPs), termed the ZBTB MBPs, that mediate transcription by specifically targeting both methylated and sequence-specific non-methylated DNA sites. DNA methylation is an essential epigenetic modification in eukaryotes required for genomic stability and control of gene expression. Consequently, aberrant alterations in genomic methyl-CpG patterns are associated with a multitude of diseases. Thus, understanding the mechanisms by which MBPs mediate inappropriate transcription is an essential component required for discerning disease onset and progression. Here we utilize an interdisciplinary approach to begin delineating the mechanisms by which this family of MBPs recognize DNA and regulate transcription. Specifically, we have designed multiple protein constructs around the zinc finger domains of the ZBTB MBPs. Biophysical characterization of the binding interactions of these protein regions with their cognate DNA targets has provided preliminary understanding for the differential mode of DNA recognition exhibited by each ZBTB MBP family members. Further, for the founding member of this family we utilized genome-wide methodologies to identify its genomic targets and the transcriptional effect of this protein in cells. Together, these findings provide initial insights into the molecular basis by which these proteins preferentially recognize, interpret and translate DNA signals into cellular transcriptional responses.

Investigating zinc finger recognition of epigenetically modified DNA

251st National Meeting (2016)

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