3.3-Å resolution cryo-EM structure of human ribonucleotide reductase with substrate and allosteric regulators bound

Edward J. Brignole, Kuang Lei Tsai, Johnathan Chittuluru, Haoran Li, Yimon Aye, Pawel A. Penczek, Jo Anne Stubbe, Catherine L. Drennan, Francisco Asturias

Research output: Contribution to journalArticle

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Abstract

Ribonucleotide reductases (RNRs) convert ribonucleotides into deoxyribonucleotides, a reaction essential for DNA replication and repair. Human RNR requires two subunits for activity, the α subunit contains the active site, and the β subunit houses the radical cofactor. Here, we present a 3.3-Å resolution structure by cryo-electron microscopy (EM) of a dATP-inhibited state of human RNR. This structure, which was determined in the presence of substrate CDP and allosteric regulators ATP and dATP, has three α2 units arranged in an α6 ring. At near-atomic resolution, these data provide insight into the molecular basis for CDP recognition by allosteric specificity effectors dATP/ATP. Additionally, we present lower-resolution EM structures of human α6 in the presence of both the anticancer drug clofarabine triphosphate and β2. Together, these structures support a model for RNR inhibition in which β2 is excluded from binding in a radical transfer competent position when α exists as a stable hexamer.

LanguageEnglish
Article numbere31502
JournaleLife
Volume7
DOIs
StatePublished - Feb 20 2018

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Cryoelectron Microscopy
Ribonucleotide Reductases
Cytidine Diphosphate
Adenosine Triphosphate
Deoxyribonucleotides
Ribonucleotides
DNA Replication
DNA Repair
Catalytic Domain
Electron Microscopy
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Brignole, E. J., Tsai, K. L., Chittuluru, J., Li, H., Aye, Y., Penczek, P. A., ... Asturias, F. (2018). 3.3-Å resolution cryo-EM structure of human ribonucleotide reductase with substrate and allosteric regulators bound. eLife, 7, [e31502]. https://doi.org/10.7554/eLife.31502

3.3-Å resolution cryo-EM structure of human ribonucleotide reductase with substrate and allosteric regulators bound. / Brignole, Edward J.; Tsai, Kuang Lei; Chittuluru, Johnathan; Li, Haoran; Aye, Yimon; Penczek, Pawel A.; Stubbe, Jo Anne; Drennan, Catherine L.; Asturias, Francisco.

In: eLife, Vol. 7, e31502, 20.02.2018.

Research output: Contribution to journalArticle

Brignole, EJ, Tsai, KL, Chittuluru, J, Li, H, Aye, Y, Penczek, PA, Stubbe, JA, Drennan, CL & Asturias, F 2018, '3.3-Å resolution cryo-EM structure of human ribonucleotide reductase with substrate and allosteric regulators bound' eLife, vol. 7, e31502. https://doi.org/10.7554/eLife.31502
Brignole, Edward J. ; Tsai, Kuang Lei ; Chittuluru, Johnathan ; Li, Haoran ; Aye, Yimon ; Penczek, Pawel A. ; Stubbe, Jo Anne ; Drennan, Catherine L. ; Asturias, Francisco. / 3.3-Å resolution cryo-EM structure of human ribonucleotide reductase with substrate and allosteric regulators bound. In: eLife. 2018 ; Vol. 7.
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