DNA wrapping and distortion by an oligomeric homeodomain protein

Williams, Hannah; Jayaraman, Padma-Sheela; Gaston, Kevin

Abstract

Many transcription factors alter DNA or chromatin structure. Changes in chromatin structure are often brought about by the recruitment of chromatin-binding proteins, chromatin-modifying proteins, or other transcription co-activator or co-repressor proteins. However, some transcription factors form oligomeric assemblies that may themselves induce changes in DNA conformation and chromatin structure. The proline-rich homeodomain (PRH/Hex) protein is a transcription factor that regulates cell differentiation and cell proliferation, and has multiple roles in embryonic development. Earlier, we showed that PRH can repress transcription by multiple mechanisms, including the recruitment of co-repressor proteins belonging to the TLE family of chromatin-binding proteins. Our in vivo crosslinking studies have shown that PRH forms oligomeric complexes in cells and a variety of biophysical techniques suggest that the protein forms octamers. However, as yet we have little knowledge of the role played by PRH oligomerisation in the regulation of promoter activity or of the architecture of promoters that are regulated directly by PRH in cells. Here, we compare the binding of PRH and the isolated PRH homeodomain to DNA fragments with single and multiple PRH sites, using gel retardation assays and DNase I and chemical footprinting. We show that the PRH oligomer binds to multiple sites within the human Goosecoid promoter with high affinity and that the binding of PRH brings about DNA distortion. We suggest that PRH octamers wrap DNA in order to bring about transcriptional repression.

• Journal Article Type: Article
• Acceptance Date: Aug 1, 2008
• Online Publication Date: Aug 7, 2008
• Publication Date: Oct 31, 2008
• Deposit Date: Nov 1, 2018
• Journal: Journal of Molecular Biology
• Print ISSN: 0022-2836
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 383
• Pages: 10-23
• DOI: https://doi.org/10.1016/j.jmb.2008.08.004
• Keywords: transcription; oligomerisation; chromatin; PRH/Hex; Goosecoid
• Public URL: https://nottingham-repository.worktribe.com/output/1037697
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0022283608009753