The structural basis of RNA-catalyzed RNA polymerization
Early life presumably required polymerase ribozymes capable of replicating RNA. Known polymerase ribozymes best approximating such replicases use as their catalytic engine an RNA-ligase ribozyme originally selected from random RNA sequences. Here we report 3.15-Å crystal structures of this ligase tr...
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| Format: | Article |
| Language: | en_US |
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Elsevier B.V.
2014
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| Online Access: | http://hdl.handle.net/1721.1/83621 https://orcid.org/0000-0002-3872-2856 |
| _version_ | 1826199095118659584 |
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| author | Shechner, David M Bartel, David Shechner, David M. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Shechner, David M Bartel, David Shechner, David M. |
| author_sort | Shechner, David M |
| collection | MIT |
| description | Early life presumably required polymerase ribozymes capable of replicating RNA. Known polymerase ribozymes best approximating such replicases use as their catalytic engine an RNA-ligase ribozyme originally selected from random RNA sequences. Here we report 3.15-Å crystal structures of this ligase trapped in catalytically viable preligation states, with the 3′-hydroxyl nucleophile positioned for in-line attack on the 5′-triphosphate. Guided by metal- and solvent-mediated interactions, the 5′-triphosphate hooks into the major groove of the adjoining RNA duplex in an unanticipated conformation. Two phosphates and the nucleophile jointly coordinate an active-site metal ion. Atomic mutagenesis experiments demonstrate that active-site nucleobase and hydroxyl groups also participate directly in catalysis, collectively playing a role that in proteinaceous polymerases is performed by a second metal ion. Thus artificial ribozymes can use complex catalytic strategies that differ markedly from those of analogous biological enzymes. |
| first_indexed | 2024-09-23T11:14:40Z |
| format | Article |
| id | mit-1721.1/83621 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:14:40Z |
| publishDate | 2014 |
| publisher | Elsevier B.V. |
| record_format | dspace |
| spelling | mit-1721.1/836212022-10-01T02:16:48Z The structural basis of RNA-catalyzed RNA polymerization Shechner, David M Bartel, David Shechner, David M. Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Bartel, David Shechner, David M. Early life presumably required polymerase ribozymes capable of replicating RNA. Known polymerase ribozymes best approximating such replicases use as their catalytic engine an RNA-ligase ribozyme originally selected from random RNA sequences. Here we report 3.15-Å crystal structures of this ligase trapped in catalytically viable preligation states, with the 3′-hydroxyl nucleophile positioned for in-line attack on the 5′-triphosphate. Guided by metal- and solvent-mediated interactions, the 5′-triphosphate hooks into the major groove of the adjoining RNA duplex in an unanticipated conformation. Two phosphates and the nucleophile jointly coordinate an active-site metal ion. Atomic mutagenesis experiments demonstrate that active-site nucleobase and hydroxyl groups also participate directly in catalysis, collectively playing a role that in proteinaceous polymerases is performed by a second metal ion. Thus artificial ribozymes can use complex catalytic strategies that differ markedly from those of analogous biological enzymes. National Institutes of Health (U.S.) (grant GM06183) National Center for Research Resources (U.S.) (award RR-1530) United States. Dept. of Energy (Office of Basic Energy Sciences, under Contract No. DE-AC02-06CH1135) 2014-01-08T21:33:44Z 2014-01-08T21:33:44Z 2011-08 2011-01 Article http://purl.org/eprint/type/JournalArticle 1545-9993 1545-9985 http://hdl.handle.net/1721.1/83621 Shechner, David M, and David P Bartel. “The structural basis of RNA-catalyzed RNA polymerization.” Nature Structural & Molecular Biology 18, no. 9 (August 21, 2011): 1036-1042. https://orcid.org/0000-0002-3872-2856 en_US http://dx.doi.org/10.1038/nsmb.2107 Nature Structural & Molecular Biology Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf Elsevier B.V. PMC |
| spellingShingle | Shechner, David M Bartel, David Shechner, David M. The structural basis of RNA-catalyzed RNA polymerization |
| title | The structural basis of RNA-catalyzed RNA polymerization |
| title_full | The structural basis of RNA-catalyzed RNA polymerization |
| title_fullStr | The structural basis of RNA-catalyzed RNA polymerization |
| title_full_unstemmed | The structural basis of RNA-catalyzed RNA polymerization |
| title_short | The structural basis of RNA-catalyzed RNA polymerization |
| title_sort | structural basis of rna catalyzed rna polymerization |
| url | http://hdl.handle.net/1721.1/83621 https://orcid.org/0000-0002-3872-2856 |
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