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• W2022028319 endingPage "1872" @default.
• W2022028319 startingPage "1857" @default.
• W2022028319 abstract "Flap endonuclease 1 (Fen1) is a highly conserved structure-specific nuclease that catalyses a specific incision to remove 5' flaps in double-stranded DNA substrates. Fen1 plays an essential role in key cellular processes, such as DNA replication and repair, and mutations that compromise Fen1 expression levels or activity have severe health implications in humans. The nuclease activity of Fen1 and other FEN family members can be stimulated by processivity clamps such as proliferating cell nuclear antigen (PCNA); however, the exact mechanism of PCNA activation is currently unknown. Here, we have used a combination of ensemble and single-molecule Förster resonance energy transfer together with protein-induced fluorescence enhancement to uncouple and investigate the substrate recognition and catalytic steps of Fen1 and Fen1/PCNA complexes. We propose a model in which upon Fen1 binding, a highly dynamic substrate is bent and locked into an open flap conformation where specific Fen1/DNA interactions can be established. PCNA enhances Fen1 recognition of the DNA substrate by further promoting the open flap conformation in a step that may involve facilitated threading of the 5' ssDNA flap. Merging our data with existing crystallographic and molecular dynamics simulations we provide a solution-based model for the Fen1/PCNA/DNA ternary complex." @default.
• W2022028319 created "2016-06-24" @default.
• W2022028319 creator A5016290104 @default.
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• W2022028319 creator A5072112447 @default.
• W2022028319 creator A5057776872 @default.
• W2022028319 date "2013-11-13" @default.
• W2022028319 modified "2023-10-17" @default.
• W2022028319 title "Single-molecule characterization of Fen1 and Fen1/PCNA complexes acting on flap substrates" @default.
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• W2022028319 doi "https://doi.org/10.1093/nar/gkt1116" @default.
• W2022028319 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3919604" @default.
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• W2022028319 hasPublicationYear "2013" @default.
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