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- W2031275732 abstract "Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DCRecognition of DNA damage by repair proteins is crucial for genomic stability. Several repair enzymes recognize aberrant DNA structural features, including nicks, gaps, and abasic sites, in a sequence-independent DNA manner. Reported here is a strategy to model these protein-DNA interactions using small proteins. A phage-displayed library of approximately one hundred million distinct zinc fingers was engineered at the DNA level based on the DNA binding domain of the repair protein poly(ADP-ribose) polymerase. Eight codons, representing amino acid residues in ‘loop’ regions of the zinc finger structure, were randomized using an NNK codon scheme. The DNA cassette was cloned into a commercially obtained T7 phagemid vector. The resultant phage library was exposed to nicked DNA molecules, which were attached to magnetic beads via a biotin-streptavidin linkage. In four rounds of selection that featured negative selection against non-nicked DNA and streptavidin coated beads, viral retention on the affinity resin increased from a negligible quantity to 62%. DNA sequencing following the final round revealed near convergence to a single, 39-mer peptide. This selected peptide will be generated by bacterial expression and assayed for binding activity using multiple strategies. While phage display of sequence specific DNA binding proteins is well-precedented, selection of peptides that bind specific DNA structures has not been reported. Molecules discovered by this strategy will help illuminate the structure-function relationships of proteins that recognize distinct DNA structures and may lead to new anticancer approaches.Citation Format: Rachel M. Guerra, Kristina M. Langenborg, Kayla M. Gross, Emily V. Sher, John W. Gilboy, Dylan Plaskon, Kevin P. Rice. A selection strategy for the design of small proteins that can distinguish DNA repair intermediates. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2236. doi:10.1158/1538-7445.AM2013-2236" @default.
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- W2031275732 date "2013-04-15" @default.
- W2031275732 modified "2023-09-25" @default.
- W2031275732 title "Abstract 2236: A selection strategy for the design of small proteins that can distinguish DNA repair intermediates." @default.
- W2031275732 doi "https://doi.org/10.1158/1538-7445.am2013-2236" @default.
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