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dc.contributor.authorMiles, Zachary D.
dc.contributor.authorMaiocco, Stephanie J.
dc.contributor.authorElliott, Sean J.
dc.contributor.authorBandarian, Vahe
dc.contributor.authorDowling, Daniel P.
dc.contributor.authorKoehrer, Caroline
dc.contributor.authorDrennan, Catherine L.
dc.date.accessioned2017-03-23T19:29:13Z
dc.date.available2017-03-23T19:29:13Z
dc.date.issued2016-09
dc.date.submitted2016-08
dc.identifier.issn0305-1048
dc.identifier.issn1362-4962
dc.identifier.urihttp://hdl.handle.net/1721.1/107678
dc.description.abstractQueuosine (Q) was discovered in the wobble position of a transfer RNA (tRNA) 47 years ago, yet the final biosynthetic enzyme responsible for Q-maturation, epoxyqueuosine (oQ) reductase (QueG), was only recently identified. QueG is a cobalamin (Cbl)-dependent, [4Fe-4S] cluster-containing protein that produces the hypermodified nucleoside Q in situ on four tRNAs. To understand how QueG is able to perform epoxide reduction, an unprecedented reaction for a Cbl-dependent enzyme, we have determined a series of high resolution structures of QueG from Bacillus subtilis. Our structure of QueG bound to a tRNA[superscript Tyr] anticodon stem loop shows how this enzyme uses a HEAT-like domain to recognize the appropriate anticodons and position the hypermodified nucleoside into the enzyme active site. We find Q bound directly above the Cbl, consistent with a reaction mechanism that involves the formation of a covalent Cbl-tRNA intermediate. Using protein film electrochemistry, we show that two [4Fe-4S] clusters adjacent to the Cbl have redox potentials in the range expected for Cbl reduction, suggesting how Cbl can be activated for nucleophilic attack on oQ. Together, these structural and electrochemical data inform our understanding of Cbl dependent nucleic acid modification.en_US
dc.description.sponsorshipNational Science Foundation (U.S.) (MCB 1122977)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (GM72623 S01, GM120283, and GM17151)en_US
dc.language.isoen_US
dc.publisherOxford University Pressen_US
dc.relation.isversionofhttp://dx.doi.org/10.1093/nar/gkw806en_US
dc.rightsCreative Commons Attribution-NonCommercial 4.0 Internationalen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/en_US
dc.sourceOxford University Pressen_US
dc.titleMolecular basis of cobalamin-dependent RNA modificationen_US
dc.typeArticleen_US
dc.identifier.citationDowling, Daniel P. et al. “Molecular Basis of Cobalamin-Dependent RNA Modification.” Nucleic Acids Research (2016): gkw806.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biologyen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemistryen_US
dc.contributor.mitauthorDowling, Daniel P.
dc.contributor.mitauthorKoehrer, Caroline
dc.contributor.mitauthorDrennan, Catherine L.
dc.relation.journalNucleic Acids Researchen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsDowling, Daniel P.; Miles, Zachary D.; Köhrer, Caroline; Maiocco, Stephanie J.; Elliott, Sean J.; Bandarian, Vahe; Drennan, Catherine L.en_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0001-5486-2755
dspace.mitauthor.errortrue
mit.licensePUBLISHER_CCen_US


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