Hesketh, Emma L, Parker-Manuel, Richard P, Chaban, Yuriy et al. (4 more authors) (2015) DNA Induces Conformational Changes in a Recombinant Human Minichromosome Maintenance Complex. The Journal of biological chemistry. 622738. ISSN 1083-351X
Abstract
ATP-dependent DNA unwinding activity has been demonstrated for recombinant archaeal homohexameric minichromosome maintenance (MCM) complexes and their yeast heterohexameric counterparts, but in higher eukaryotes such as Drosophila, MCM-associated DNA helicase activity has only been observed in the context of a co-purified Cdc45-MCM-GINS (CMG) complex. Here we describe the production of recombinant human MCM complex (hMCM) in E. coli. This protein displays ATP hydrolysis activity and is capable of unwinding duplex DNA. Using single particle asymmetric electron microscopy reconstruction, we demonstrate recombinant hMCM forms a hexamer that undergoes a conformational change when bound to DNA. Recombinant hMCM produced without post-translational modifications is functional in vitro and provides an important tool for the biochemical reconstitution of the human replicative helicase.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2015, The American Society for Biochemistry and Molecular Biology, Inc. This is an Open Access article made available under a Creative Commons Attribution licence which allows sharing and reuse provided that the authors and source are properly acknowledged. |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) |
Depositing User: | Pure (York) |
Date Deposited: | 05 Jun 2015 23:01 |
Last Modified: | 21 Jan 2025 17:17 |
Published Version: | https://doi.org/10.1074/jbc.M114.622738 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1074/jbc.M114.622738 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:86835 |
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Description: DNA Induces Conformational Changes in a Recombinant Human Minichromosome Maintenance Complex