Protein flexibility directs DNA recognition by the papillomavirus E2 proteins

Brown, Craig A.; Campos-Le�n, Karen; Strickland, Maddy; Williams, Christopher; Fairweather, Victoria; Brady, R. Leo; Crump, Matthew P.; Gaston, Kevin

Abstract

Although DNA flexibility is known to play an important role in DNA–protein interactions, the importance of protein flexibility is less well understood. Here, we show that protein dynamics are important in DNA recognition using the well-characterized human papillomavirus (HPV) type 6 E2 protein as a model system. We have compared the DNA binding properties of the HPV 6 E2 DNA binding domain (DBD) and a mutant lacking two C-terminal leucine residues that form part of the hydrophobic core of the protein. Deletion of these residues results in increased specific and non-specific DNA binding and an overall decrease in DNA binding specificity. Using 15N NMR relaxation and hydrogen/deuterium exchange, we demonstrate that the mutation results in increased flexibility within the hydrophobic core and loop regions that orient the DNA binding helices. Stopped-flow kinetic studies indicate that increased flexibility alters DNA binding by increasing initial interactions with DNA but has little or no effect on the structural rearrangements that follow this step. Taken together these data demonstrate that subtle changes in protein dynamics have a major influence on protein–DNA interactions.

Citation

Brown, C. A., Campos-León, K., Strickland, M., Williams, C., Fairweather, V., Brady, R. L., …Gaston, K. (2011). Protein flexibility directs DNA recognition by the papillomavirus E2 proteins. Nucleic Acids Research, 39(7), 2969--2980. https://doi.org/10.1093/nar/gkq1217

• Journal Article Type: Article
• Acceptance Date: Nov 10, 2010
• Online Publication Date: Dec 3, 2010
• Publication Date: Apr 1, 2011
• Deposit Date: Oct 31, 2018
• Publicly Available Date: Oct 31, 2018
• Journal: Nucleic Acids Research
• Print ISSN: 0305-1048
• Publisher: Oxford University Press
• Peer Reviewed: Peer Reviewed
• Volume: 39
• Issue: 7
• Pages: 2969--2980
• DOI: https://doi.org/10.1093/nar/gkq1217
• Public URL: https://nottingham-repository.worktribe.com/output/1037623
• Publisher URL: https://academic.oup.com/nar/article/39/7/2969/1142267