Observing one-divalent-metal-ion-dependent and histidine-promoted His-Me family I-PpoI nuclease catalysis in crystallo

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dc.citation.articleNumberRP99960en_US
dc.citation.journalTitleeLifeen_US
dc.citation.volumeNumber13en_US
dc.contributor.authorChang, Caleben_US
dc.contributor.authorZhou, Graceen_US
dc.contributor.authorGao, Yangen_US
dc.date.accessioned2024-08-29T21:11:48Zen_US
dc.date.available2024-08-29T21:11:48Zen_US
dc.date.issued2024en_US
dc.description.abstractMetal-ion-dependent nucleases play crucial roles in cellular defense and biotechnological applications. Time-resolved crystallography has resolved catalytic details of metal-ion-dependent DNA hydrolysis and synthesis, uncovering the essential roles of multiple metal ions during catalysis. The histidine-metal (His-Me) superfamily nucleases are renowned for binding one divalent metal ion and requiring a conserved histidine to promote catalysis. Many His-Me family nucleases, including homing endonucleases and Cas9 nuclease, have been adapted for biotechnological and biomedical applications. However, it remains unclear how the single metal ion in His-Me nucleases, together with the histidine, promotes water deprotonation, nucleophilic attack, and phosphodiester bond breakage. By observing DNA hydrolysis in crystallo with His-Me I-PpoI nuclease as a model system, we proved that only one divalent metal ion is required during its catalysis. Moreover, we uncovered several possible deprotonation pathways for the nucleophilic water. Interestingly, binding of the single metal ion and water deprotonation are concerted during catalysis. Our results reveal catalytic details of His-Me nucleases, which is distinct from multi-metal-ion-dependent DNA polymerases and nucleases.en_US
dc.identifier.citationChang, C., Zhou, G., & Gao, Y. (2024). Observing one-divalent-metal-ion-dependent and histidine-promoted His-Me family I-PpoI nuclease catalysis in crystallo. eLife, 13, RP99960. https://doi.org/10.7554/eLife.99960en_US
dc.identifier.digitalelife-99960-v1en_US
dc.identifier.doihttps://doi.org/10.7554/eLife.99960.3en_US
dc.identifier.urihttps://hdl.handle.net/1911/117733en_US
dc.language.isoengen_US
dc.publishereLife Sciences Publications Ltden_US
dc.rightsExcept where otherwise noted, this work is licensed under a Creative Commons Attribution (CC BY) license.  Permission to reuse, publish, or reproduce the work beyond the terms of the license or beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.en_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en_US
dc.titleObserving one-divalent-metal-ion-dependent and histidine-promoted His-Me family I-PpoI nuclease catalysis in crystalloen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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