Molecular basis of cobalamin-dependent RNA modification
Queuosine (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...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Oxford University Press
2017
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| Online Access: | http://hdl.handle.net/1721.1/107678 https://orcid.org/0000-0001-5486-2755 |
| _version_ | 1826213264536633344 |
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| author | Miles, Zachary D. Maiocco, Stephanie J. Elliott, Sean J. Bandarian, Vahe Dowling, Daniel P. Koehrer, Caroline Drennan, Catherine L. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Miles, Zachary D. Maiocco, Stephanie J. Elliott, Sean J. Bandarian, Vahe Dowling, Daniel P. Koehrer, Caroline Drennan, Catherine L. |
| author_sort | Miles, Zachary D. |
| collection | MIT |
| description | Queuosine (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. |
| first_indexed | 2024-09-23T15:46:38Z |
| format | Article |
| id | mit-1721.1/107678 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:46:38Z |
| publishDate | 2017 |
| publisher | Oxford University Press |
| record_format | dspace |
| spelling | mit-1721.1/1076782022-10-02T04:01:15Z Molecular basis of cobalamin-dependent RNA modification Miles, Zachary D. Maiocco, Stephanie J. Elliott, Sean J. Bandarian, Vahe Dowling, Daniel P. Koehrer, Caroline Drennan, Catherine L. Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Dowling, Daniel P. Koehrer, Caroline Drennan, Catherine L. Queuosine (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. National Science Foundation (U.S.) (MCB 1122977) National Institutes of Health (U.S.) (GM72623 S01, GM120283, and GM17151) 2017-03-23T19:29:13Z 2017-03-23T19:29:13Z 2016-09 2016-08 Article http://purl.org/eprint/type/JournalArticle 0305-1048 1362-4962 http://hdl.handle.net/1721.1/107678 Dowling, Daniel P. et al. “Molecular Basis of Cobalamin-Dependent RNA Modification.” Nucleic Acids Research (2016): gkw806. https://orcid.org/0000-0001-5486-2755 en_US http://dx.doi.org/10.1093/nar/gkw806 Nucleic Acids Research Creative Commons Attribution-NonCommercial 4.0 International http://creativecommons.org/licenses/by-nc/4.0/ application/pdf Oxford University Press Oxford University Press |
| spellingShingle | Miles, Zachary D. Maiocco, Stephanie J. Elliott, Sean J. Bandarian, Vahe Dowling, Daniel P. Koehrer, Caroline Drennan, Catherine L. Molecular basis of cobalamin-dependent RNA modification |
| title | Molecular basis of cobalamin-dependent RNA modification |
| title_full | Molecular basis of cobalamin-dependent RNA modification |
| title_fullStr | Molecular basis of cobalamin-dependent RNA modification |
| title_full_unstemmed | Molecular basis of cobalamin-dependent RNA modification |
| title_short | Molecular basis of cobalamin-dependent RNA modification |
| title_sort | molecular basis of cobalamin dependent rna modification |
| url | http://hdl.handle.net/1721.1/107678 https://orcid.org/0000-0001-5486-2755 |
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