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dc.contributor.authorMandal, Debabrata
dc.contributor.authorKoehrer, Caroline
dc.contributor.authorSu, Dan
dc.contributor.authorBabu, I. Ramesh
dc.contributor.authorChan, Tsz Yan Clement
dc.contributor.authorLiu, Yuchen
dc.contributor.authorSoll, Dieter
dc.contributor.authorBlum, Paul
dc.contributor.authorKuwahara, Masayasu
dc.contributor.authorDedon, Peter C.
dc.contributor.authorRajbhandary, Uttam L.
dc.date.accessioned2014-12-02T17:26:22Z
dc.date.available2014-12-02T17:26:22Z
dc.date.issued2013-12
dc.identifier.issn1355-8382
dc.identifier.urihttp://hdl.handle.net/1721.1/91989
dc.description.abstractMost archaea and bacteria use a modified C in the anticodon wobble position of isoleucine tRNA to base pair with A but not with G of the mRNA. This allows the tRNA to read the isoleucine codon AUA without also reading the methionine codon AUG. To understand why a modified C, and not U or modified U, is used to base pair with A, we mutated the C34 in the anticodon of Haloarcula marismortui isoleucine tRNA (tRNA2Ile) to U, expressed the mutant tRNA in Haloferax volcanii, and purified and analyzed the tRNA. Ribosome binding experiments show that although the wild-type tRNA2Ile binds exclusively to the isoleucine codon AUA, the mutant tRNA binds not only to AUA but also to AUU, another isoleucine codon, and to AUG, a methionine codon. The G34 to U mutant in the anticodon of another H. marismortui isoleucine tRNA species showed similar codon binding properties. Binding of the mutant tRNA to AUG could lead to misreading of the AUG codon and insertion of isoleucine in place of methionine. This result would explain why most archaea and bacteria do not normally use U or a modified U in the anticodon wobble position of isoleucine tRNA for reading the codon AUA. Biochemical and mass spectrometric analyses of the mutant tRNAs have led to the discovery of a new modified nucleoside, 5-cyanomethyl U in the anticodon wobble position of the mutant tRNAs. 5-Cyanomethyl U is present in total tRNAs from euryarchaea but not in crenarchaea, eubacteria, or eukaryotes.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (GM17151)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (GM22854)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (ES017010)en_US
dc.description.sponsorshipSingapore-MIT Alliance for Research and Technologyen_US
dc.description.sponsorshipSingapore. National Research Foundationen_US
dc.description.sponsorshipUnited States. Dept. of Energy (DE-FG36-08GO88055)en_US
dc.language.isoen_US
dc.publisherCold Spring Harbor Laboratory Press/RNA Societyen_US
dc.relation.isversionofhttp://dx.doi.org/10.1261/rna.042358.113en_US
dc.rightsCreative Commons Attribution-Noncommericalen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc/3.0/en_US
dc.titleIdentification and codon reading properties of 5-cyanomethyl uridine, a new modified nucleoside found in the anticodon wobble position of mutant haloarchaeal isoleucine tRNAsen_US
dc.typeArticleen_US
dc.identifier.citationMandal, D., C. Kohrer, D. Su, I. R. Babu, C. T. Y. Chan, Y. Liu, D. Soll, et al. “Identification and Codon Reading Properties of 5-Cyanomethyl Uridine, a New Modified Nucleoside Found in the Anticodon Wobble Position of Mutant Haloarchaeal Isoleucine tRNAs.” RNA 20, no. 2 (December 16, 2013): 177–188.en_US
dc.contributor.departmentInstitute for Medical Engineering and Scienceen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biologyen_US
dc.contributor.mitauthorMandal, Debabrataen_US
dc.contributor.mitauthorKoehrer, Carolineen_US
dc.contributor.mitauthorSu, Danen_US
dc.contributor.mitauthorBabu, I. Rameshen_US
dc.contributor.mitauthorChan, Tsz Yan Clementen_US
dc.contributor.mitauthorDedon, Peter C.en_US
dc.contributor.mitauthorRajbhandary, Uttam L.en_US
dc.relation.journalRNAen_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.orderedauthorsMandal, D.; Kohrer, C.; Su, D.; Babu, I. R.; Chan, C. T. Y.; Liu, Y.; Soll, D.; Blum, P.; Kuwahara, M.; Dedon, P. C.; RajBhandary, U. L.en_US
dc.identifier.orcidhttps://orcid.org/0000-0003-0011-3067
dc.identifier.orcidhttps://orcid.org/0000-0002-4530-5647
dc.identifier.orcidhttps://orcid.org/0000-0001-8533-2706
dc.identifier.orcidhttps://orcid.org/0000-0001-7940-3459
dspace.mitauthor.errortrue
mit.licensePUBLISHER_CCen_US


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