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Identifying ZBTB38 Protein Interacting Partners

Semester: Summer 2023

Presentation description

Healthy cellular functioning requires tight regulation of transcriptional processes. DNA methylation at CpG sites (mCpG) represents a regulatory mechanism through which cells exert such control. DNA methylation is an abundant epigenetic modification under tight regulation within the cell. Dysregulation of DNA methylation patterns, such as hypo- or hypermethylation at CpG sites, can result in altered gene expression, which has been associated with several diseases, including cancer. At many gene promoters, DNA methylation is necessary but insufficient to result in changes to gene expression. Serving an intermediary role at these promoters, methyl-CpG binding proteins (MBPs) recognize and bind mCpG sites and recruit additional proteins that ultimately regulate transcription. Despite the integral role of MBPs in gene expression and disease pathogenesis, a comprehensive understanding of how MBP transcription factors (TFs) regulate gene expression must still be fully understood. ZBTB38 is a member of the zinc finger (ZF) BTB/POZ family of transcription factors (TFs) that selectively bind to methylated DNA. ZBTB38 differentiates itself in that it is the only known MBP that can selectively recognize two different mCpG sites using two separate sets of Cys2His2 ZFs. While the DNA binding capabilities of ZBTB38 is well studied, there is limited knowledge for its protein interacting partners. Increasing evidence suggests that the transcriptional activity of ZBTB38 may support a cancerous state. Hence, identifying its protein interacting partners is crucial for unraveling how epigenetic mechanisms affect disease progression. Herein, we present the design and molecular cloning for the production and biotinylation of ZBTB38 ZF domain constructs using E. coli. This methodology enables the identification of ZBTB38 binding partners using streptavidin-based methodologies to further decipher the molecular mechanism by which ZBTB38 regulates epigenetic-based transcriptional processes.

Presenter Name: Shritha Gummadi
Presentation Type: Poster
Presentation Format: In Person
Presentation #72
College: Science
School / Department: Chemistry
Research Mentor: Bethany Buck
Date | Time: Thursday, Aug 3rd | 10:30 AM