Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase
Essential for DNA biosynthesis and repair, ribonucleotide reductases (RNRs) convert ribonucleotides to deoxyribonucleotides via radical-based chemistry. Although long known that allosteric regulation of RNR activity is vital for cell health, the molecular basis of this regulation has been enigmatic,...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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National Academy of Sciences of the United States of America
2012
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| Online Access: | http://hdl.handle.net/1721.1/71843 https://orcid.org/0000-0001-5486-2755 https://orcid.org/0000-0001-8076-4489 |
| _version_ | 1826191938876866560 |
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| author | Ando, Nozomi Brignole, Edward J. Zimanyi, Christina M. Funk, Michael Andrew Yokoyama, Kenichi Asturias, Francisco J. Stubbe, JoAnne Drennan, Catherine L |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Ando, Nozomi Brignole, Edward J. Zimanyi, Christina M. Funk, Michael Andrew Yokoyama, Kenichi Asturias, Francisco J. Stubbe, JoAnne Drennan, Catherine L |
| author_sort | Ando, Nozomi |
| collection | MIT |
| description | Essential for DNA biosynthesis and repair, ribonucleotide reductases (RNRs) convert ribonucleotides to deoxyribonucleotides via radical-based chemistry. Although long known that allosteric regulation of RNR activity is vital for cell health, the molecular basis of this regulation has been enigmatic, largely due to a lack of structural information about how the catalytic subunit (α2) and the radical-generation subunit (β2) interact. Here we present the first structure of a complex between α2 and β2 subunits for the prototypic RNR from Escherichia coli. Using four techniques (small-angle X-ray scattering, X-ray crystallography, electron microscopy, and analytical ultracentrifugation), we describe an unprecedented α4β4 ring-like structure in the presence of the negative activity effector dATP and provide structural support for an active α2β2 configuration. We demonstrate that, under physiological conditions, E. coli RNR exists as a mixture of transient α2β2 and α4β4 species whose distributions are modulated by allosteric effectors. We further show that this interconversion between α2β2 and α4β4 entails dramatic subunit rearrangements, providing a stunning molecular explanation for the allosteric regulation of RNR activity in E. coli. |
| first_indexed | 2024-09-23T09:03:41Z |
| format | Article |
| id | mit-1721.1/71843 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:03:41Z |
| publishDate | 2012 |
| publisher | National Academy of Sciences of the United States of America |
| record_format | dspace |
| spelling | mit-1721.1/718432022-09-26T10:10:06Z Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase Ando, Nozomi Brignole, Edward J. Zimanyi, Christina M. Funk, Michael Andrew Yokoyama, Kenichi Asturias, Francisco J. Stubbe, JoAnne Drennan, Catherine L Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Drennan, Catherine L. Stubbe, JoAnne Drennan, Catherine L. Essential for DNA biosynthesis and repair, ribonucleotide reductases (RNRs) convert ribonucleotides to deoxyribonucleotides via radical-based chemistry. Although long known that allosteric regulation of RNR activity is vital for cell health, the molecular basis of this regulation has been enigmatic, largely due to a lack of structural information about how the catalytic subunit (α2) and the radical-generation subunit (β2) interact. Here we present the first structure of a complex between α2 and β2 subunits for the prototypic RNR from Escherichia coli. Using four techniques (small-angle X-ray scattering, X-ray crystallography, electron microscopy, and analytical ultracentrifugation), we describe an unprecedented α4β4 ring-like structure in the presence of the negative activity effector dATP and provide structural support for an active α2β2 configuration. We demonstrate that, under physiological conditions, E. coli RNR exists as a mixture of transient α2β2 and α4β4 species whose distributions are modulated by allosteric effectors. We further show that this interconversion between α2β2 and α4β4 entails dramatic subunit rearrangements, providing a stunning molecular explanation for the allosteric regulation of RNR activity in E. coli. National Natural Science Foundation (NIH, NIGMS) (Award DMR-0936384) National Center for Research Resources (U.S.) (NCRR) (Award RR-01646) National Institutes of Health (U.S.) (NCRR P41 program) (RR017573) National Institutes of Health (U.S.) (Grant GM29595) 2012-07-26T18:04:30Z 2012-07-26T18:04:30Z 2011-12 2011-08 Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/71843 Ando, N. et al. “Structural Interconversions Modulate Activity of Escherichia Coli Ribonucleotide Reductase.” Proceedings of the National Academy of Sciences 108.52 (2011): 21046–21051. Copyright ©2011 by the National Academy of Sciences https://orcid.org/0000-0001-5486-2755 https://orcid.org/0000-0001-8076-4489 en_US http://dx.doi.org/10.1073/pnas.1112715108 Proceedings of the National Academy of Sciences of the United States of America Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf National Academy of Sciences of the United States of America PNAS |
| spellingShingle | Ando, Nozomi Brignole, Edward J. Zimanyi, Christina M. Funk, Michael Andrew Yokoyama, Kenichi Asturias, Francisco J. Stubbe, JoAnne Drennan, Catherine L Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase |
| title | Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase |
| title_full | Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase |
| title_fullStr | Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase |
| title_full_unstemmed | Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase |
| title_short | Structural interconversions modulate activity of Escherichia coli ribonucleotide reductase |
| title_sort | structural interconversions modulate activity of escherichia coli ribonucleotide reductase |
| url | http://hdl.handle.net/1721.1/71843 https://orcid.org/0000-0001-5486-2755 https://orcid.org/0000-0001-8076-4489 |
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