Thompson, M.J., Gotham, V., Ciani, B. orcid.org/0000-0001-7223-4154 et al. (1 more author) (2018) A conserved loop-wedge motif moderates reaction site search and recognition by FEN1. Nucleic Acids Research, 46 (15). pp. 7858-7872. ISSN 0305-1048
Abstract
DNA replication and repair frequently involve intermediate two-way junction structures with overhangs, or flaps, that must be promptly removed; a task performed by the essential enzyme flap endonuclease 1 (FEN1). We demonstrate a functional relationship between two intrinsically disordered regions of the FEN1 protein, which recognise opposing sides of the junction and order in response to the requisite substrate. Our results inform a model in which short-range translocation of FEN1 on DNA facilitates search for the annealed 3′‑terminus of a primer strand, which is recognised by breaking the terminal base pair to generate a substrate with a single nucleotide 3′‑flap. This recognition event allosterically signals hydrolytic removal of the 5′-flap through reaction in the opposing junction duplex, by controlling access of the scissile phosphate diester to the active site. The recognition process relies on a highly-conserved ‘wedge’ residue located on a mobile loop that orders to bind the newly-unpaired base. The unanticipated ‘loop–wedge’ mechanism exerts control over substrate selection, rate of reaction and reaction site precision, and shares features with other enzymes that recognise irregular DNA structures. These new findings reveal how FEN1 precisely couples 3′-flap verification to function.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > Department of Chemistry (Sheffield) |
Funding Information: | Funder Grant number BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL (BBSRC) BB/M00404X/1 BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL (BBSRC) BB/J00300X/1 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 19 Jun 2018 11:12 |
Last Modified: | 19 Oct 2021 09:19 |
Status: | Published |
Publisher: | Oxford University Press |
Refereed: | Yes |
Identification Number: | 10.1093/nar/gky506 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:131906 |