Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical
All cells obtain 2′-deoxyribonucleotides for DNA synthesis through the activity of a ribonucleotide reductase (RNR). The class I RNRs found in humans and pathogenic bacteria differ in (i) use of Fe(II), Mn(II), or both for activation of the dinuclear-metallocofactor subunit, β; (ii) reaction of the...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
National Academy of Sciences
2020
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| Online Access: | https://hdl.handle.net/1721.1/125856 |
| _version_ | 1811087965138452480 |
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| author | Blaesi, Elizabeth J. Palowitch, Gavin M. Hu, Kai Kim, Amelia J. Rose, Hannah R. Alapati, Rahul Lougee, Marshall G. Kim, Hee Jong Taguchi, Alexander T Tan, Kong Ooi Laremore, Tatiana N. Griffin, Robert Guy Krebs, Carsten Matthews, Megan L. Silakov, Alexey Bollinger, J. Martin Allen, Benjamin D. Boal, Amie K. |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Blaesi, Elizabeth J. Palowitch, Gavin M. Hu, Kai Kim, Amelia J. Rose, Hannah R. Alapati, Rahul Lougee, Marshall G. Kim, Hee Jong Taguchi, Alexander T Tan, Kong Ooi Laremore, Tatiana N. Griffin, Robert Guy Krebs, Carsten Matthews, Megan L. Silakov, Alexey Bollinger, J. Martin Allen, Benjamin D. Boal, Amie K. |
| author_sort | Blaesi, Elizabeth J. |
| collection | MIT |
| description | All cells obtain 2′-deoxyribonucleotides for DNA synthesis through the activity of a ribonucleotide reductase (RNR). The class I RNRs found in humans and pathogenic bacteria differ in (i) use of Fe(II), Mn(II), or both for activation of the dinuclear-metallocofactor subunit, β; (ii) reaction of the reduced dimetal center with dioxygen or superoxide for this activation; (iii) requirement (or lack thereof) for a flavoprotein activase, NrdI, to provide the superoxide from O2; and (iv) use of either a stable tyrosyl radical or a high-valent dimetal cluster to initiate each turnover by oxidizing a cysteine residue in the α subunit to a radical (Cys•). The use of manganese by bacterial class I, subclass b-d RNRs, which contrasts with the exclusive use of iron by the eukaryotic Ia enzymes, appears to be a countermeasure of certain pathogens against iron deprivation imposed by their hosts. Here, we report a metal-free type of class I RNR (subclass e) from two human pathogens. The Cys• in its α subunit is generated by a stable, tyrosine-derived dihydroxyphenylalanine radical (DOPA•) in β. The three-electron oxidation producing DOPA• occurs in Escherichia coli only if the β is coexpressed with the NrdI activase encoded adjacently in the pathogen genome. The independence of this new RNR from transition metals, or the requirement for a single metal ion only transiently for activation, may afford the pathogens an even more potent countermeasure against transition metal-directed innate immunity. |
| first_indexed | 2024-09-23T13:54:38Z |
| format | Article |
| id | mit-1721.1/125856 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:54:38Z |
| publishDate | 2020 |
| publisher | National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/1258562022-10-01T17:56:49Z Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical Blaesi, Elizabeth J. Palowitch, Gavin M. Hu, Kai Kim, Amelia J. Rose, Hannah R. Alapati, Rahul Lougee, Marshall G. Kim, Hee Jong Taguchi, Alexander T Tan, Kong Ooi Laremore, Tatiana N. Griffin, Robert Guy Krebs, Carsten Matthews, Megan L. Silakov, Alexey Bollinger, J. Martin Allen, Benjamin D. Boal, Amie K. Massachusetts Institute of Technology. Department of Chemistry Francis Bitter Magnet Laboratory (Massachusetts Institute of Technology) All cells obtain 2′-deoxyribonucleotides for DNA synthesis through the activity of a ribonucleotide reductase (RNR). The class I RNRs found in humans and pathogenic bacteria differ in (i) use of Fe(II), Mn(II), or both for activation of the dinuclear-metallocofactor subunit, β; (ii) reaction of the reduced dimetal center with dioxygen or superoxide for this activation; (iii) requirement (or lack thereof) for a flavoprotein activase, NrdI, to provide the superoxide from O2; and (iv) use of either a stable tyrosyl radical or a high-valent dimetal cluster to initiate each turnover by oxidizing a cysteine residue in the α subunit to a radical (Cys•). The use of manganese by bacterial class I, subclass b-d RNRs, which contrasts with the exclusive use of iron by the eukaryotic Ia enzymes, appears to be a countermeasure of certain pathogens against iron deprivation imposed by their hosts. Here, we report a metal-free type of class I RNR (subclass e) from two human pathogens. The Cys• in its α subunit is generated by a stable, tyrosine-derived dihydroxyphenylalanine radical (DOPA•) in β. The three-electron oxidation producing DOPA• occurs in Escherichia coli only if the β is coexpressed with the NrdI activase encoded adjacently in the pathogen genome. The independence of this new RNR from transition metals, or the requirement for a single metal ion only transiently for activation, may afford the pathogens an even more potent countermeasure against transition metal-directed innate immunity. National Institutes of Health (Grant GM119707) 2020-06-17T20:36:29Z 2020-06-17T20:36:29Z 2018-09 2018-07 2019-12-18T13:45:33Z Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 https://hdl.handle.net/1721.1/125856 Blaesi, Elizabeth J. et al. "Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical." Proceedings of the National Academy of Sciences 115, 40 (September 2018): 10022-10027 © 2018 National Academy of Sciences en http://dx.doi.org/10.1073/pnas.1811993115 Proceedings of the National Academy of Sciences 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 PNAS |
| spellingShingle | Blaesi, Elizabeth J. Palowitch, Gavin M. Hu, Kai Kim, Amelia J. Rose, Hannah R. Alapati, Rahul Lougee, Marshall G. Kim, Hee Jong Taguchi, Alexander T Tan, Kong Ooi Laremore, Tatiana N. Griffin, Robert Guy Krebs, Carsten Matthews, Megan L. Silakov, Alexey Bollinger, J. Martin Allen, Benjamin D. Boal, Amie K. Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical |
| title | Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical |
| title_full | Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical |
| title_fullStr | Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical |
| title_full_unstemmed | Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical |
| title_short | Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine-derived dihydroxyphenylalanine radical |
| title_sort | metal free class ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosine derived dihydroxyphenylalanine radical |
| url | https://hdl.handle.net/1721.1/125856 |
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