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dc.contributor.authorGaubitz, Christl
dc.contributor.authorLiu, Xingchen
dc.contributor.authorPajak, Joshua
dc.contributor.authorStone, Nicholas P
dc.contributor.authorHayes, Janelle A
dc.contributor.authorDemo, Gabriel
dc.contributor.authorKelch, Brian A
dc.date.accessioned2023-07-11T14:08:06Z
dc.date.available2023-07-11T14:08:06Z
dc.date.issued2022-02-18
dc.identifier.citationGaubitz C, Liu X, Pajak J, Stone NP, Hayes JA, Demo G, Kelch BA. Cryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loader. Elife. 2022 Feb 18;11:e74175. doi: 10.7554/eLife.74175. PMID: 35179493; PMCID: PMC8893722.en_US
dc.identifier.eissn2050-084X
dc.identifier.doi10.7554/eLife.74175en_US
dc.identifier.pmid35179493
dc.identifier.urihttp://hdl.handle.net/20.500.14038/52268
dc.description.abstractSliding clamps are ring-shaped protein complexes that are integral to the DNA replication machinery of all life. Sliding clamps are opened and installed onto DNA by clamp loader AAA+ ATPase complexes. However, how a clamp loader opens and closes the sliding clamp around DNA is still unknown. Here, we describe structures of the Saccharomyces cerevisiae clamp loader Replication Factor C (RFC) bound to its cognate sliding clamp Proliferating Cell Nuclear Antigen (PCNA) en route to successful loading. RFC first binds to PCNA in a dynamic, closed conformation that blocks both ATPase activity and DNA binding. RFC then opens the PCNA ring through a large-scale 'crab-claw' expansion of both RFC and PCNA that explains how RFC prefers initial binding of PCNA over DNA. Next, the open RFC:PCNA complex binds DNA and interrogates the primer-template junction using a surprising base-flipping mechanism. Our structures indicate that initial PCNA opening and subsequent closure around DNA do not require ATP hydrolysis, but are driven by binding energy. ATP hydrolysis, which is necessary for RFC release, is triggered by interactions with both PCNA and DNA, explaining RFC's switch-like ATPase activity. Our work reveals how a AAA+ machine undergoes dramatic conformational changes for achieving binding preference and substrate remodeling.en_US
dc.language.isoenen_US
dc.relation.ispartofeLifeen_US
dc.relation.urlhttps://doi.org/10.7554/elife.74175en_US
dc.rightsCopyright Gaubitz et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.en_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectAAA+en_US
dc.subjectDNA replicationen_US
dc.subjectS. cerevisiaeen_US
dc.subjectmolecular biophysicsen_US
dc.subjectsliding clamp loaderen_US
dc.subjectstructural biologyen_US
dc.titleCryo-EM structures reveal high-resolution mechanism of a DNA polymerase sliding clamp loaderen_US
dc.typeJournal Articleen_US
dc.source.journaltitleeLife
dc.source.volume11
dc.source.countryUnited States
dc.source.countryEngland
dc.identifier.journaleLife
refterms.dateFOA2023-07-11T14:08:07Z
dc.contributor.departmentBiochemistry and Molecular Biotechnologyen_US
dc.contributor.departmentRNA Therapeutics Instituteen_US


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Copyright Gaubitz et al. This
article is distributed under the
terms of the Creative Commons
Attribution License, which
permits unrestricted use and
redistribution provided that the
original author and source are
credited.
Except where otherwise noted, this item's license is described as Copyright Gaubitz et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.