Since the origins of DNA-based life, the enzyme ribonucleotide reductase (RNR) has spurred proliferation because of its rate-limiting role in de novo deoxynucleoside-triphosphate (dNTP) biosynthesis. Paradoxically, the large subunit, RNR-α, of this obligatory two-component complex in mammals plays a context-specific antiproliferative role. There is little explanation for this dichotomy. Here, we show that RNR-α has a previously unrecognized DNA-replication inhibition function, leading to growth retardation. This underappreciated biological activity functions in the nucleus, where RNR-α interacts with ZRANB3. This process suppresses ZRANB3’s function in unstressed cells, which we show to promote DNA synthesis. This nonreductase function of RNR-α is promoted by RNR-α hexamerization—induced by a natural and synthetic nucleotide of dA/ClF/CLA/FLU—which elicits rapid RNR-α nuclear import. The newly discovered nuclear signaling axis is a primary defense against elevated or imbalanced dNTP pools that can exert mutagenic effects irrespective of the cell cycle.
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Thanks to members of the individual labs who generously provided plasmids and shRNAs as indicated in on-line methods; J. Page for contributing to the creation of RNR-α(D57N) knock-in mice; A. Arnaoutov (Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH) for pfastbac-GST-IRBIT; V. Gorbunova (University of Rochester) for NHEJ-reporter plasmids; J. Yuan (Columbia University) for the plasmids SFB-ZRANB3, SFB-ZRANB3-Δ PIP and SFBZRANB3(Q519A); D. Ahel (Oxford University) for the plasmids YFP-ZRANB3 and Flag-ZRANB3; Z. Zhang (University of Delaware) for the plasmid pet15b-His5-PCNA; A. Grimson (Cornell University) for the shRNA plasmids for RNR-α, RNR-β and IRBIT. Research: Pershing Square Sohn Cancer Research Alliance grant (to Y.A.); Meyer Cancer Center grant (Weill Cornell Medicine) (to Y.A. and R.S.W.); and the Canadian Institutes of Health Research grant (MOP-82930) (to J.O.). Instrumentation and shared supplies: NIH DP2 New Innovator (1DP2GM114850); NSF CAREER (CHE-1351400); Office of Naval Research (ONR) Young Investigator (N00014-17-1-2529); Beckman Young Investigator; Sloan Fellowship (FG-2016-6379) (to Y.A.); Cornell NMR facility (NSF MRI: CHE-1531632; PI: Y.A.) and Cornell Imaging Center (NIH 1S10RR025502; PI: R.M. Williams).
The authors declare no competing interests.
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Fu, Y., Long, M.J.C., Wisitpitthaya, S. et al. Nuclear RNR-α antagonizes cell proliferation by directly inhibiting ZRANB3. Nat Chem Biol 14, 943–954 (2018). https://doi.org/10.1038/s41589-018-0113-5
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