Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 10022-10027 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 115 |
| Issue number | 40 |
| DOIs | |
| State | Published - Oct 2 2018 |
Fingerprint
All Science Journal Classification (ASJC) codes
- General
Cite this
}
Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosinederived 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 G.; Krebs, Carsten; Matthews, Megan L.; Silakov, Alexey; Bollinger, J. Martin; Allen, Benjamin D.; Boal, Amie K.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, No. 40, 02.10.2018, p. 10022-10027.Research output: Contribution to journal › Article
TY - JOUR
T1 - Metal-free class Ie ribonucleotide reductase from pathogens initiates catalysis with a tyrosinederived dihydroxyphenylalanine radical
AU - Blaesi, Elizabeth J.
AU - Palowitch, Gavin M.
AU - Hu, Kai
AU - Kim, Amelia J.
AU - Rose, Hannah R.
AU - Alapati, Rahul
AU - Lougee, Marshall G.
AU - Kim, Hee Jong
AU - Taguchi, Alexander T.
AU - Tan, Kong Ooi
AU - Laremore, Tatiana N.
AU - Griffin, Robert G.
AU - Krebs, Carsten
AU - Matthews, Megan L.
AU - Silakov, Alexey
AU - Bollinger, J. Martin
AU - Allen, Benjamin D.
AU - Boal, Amie K.
PY - 2018/10/2
Y1 - 2018/10/2
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85054393678&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85054393678&partnerID=8YFLogxK
U2 - 10.1073/pnas.1811993115
DO - 10.1073/pnas.1811993115
M3 - Article
C2 - 30224458
AN - SCOPUS:85054393678
VL - 115
SP - 10022
EP - 10027
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 40
ER -