Structures of Class Id Ribonucleotide Reductase Catalytic Subunits Reveal a Minimal Architecture for Deoxynucleotide Biosynthesis

Rose, Hannah R.; Maggiolo, Ailiena O.; McBride, Molly J.; Palowitch, Gavin M.; Pandelia, Maria-Eirini; Davis, Katherine M.; Yennawar, Neela H.; Boal, Amie K.

doi:10.1021/acs.biochem.8b01252

Structures of Class Id Ribonucleotide Reductase Catalytic Subunits Reveal a Minimal Architecture for Deoxynucleotide Biosynthesis

Journal Article · Mon Mar 11 00:00:00 EDT 2019 · Biochemistry

DOI:https://doi.org/10.1021/acs.biochem.8b01252· OSTI ID:1544859

^[1]; Maggiolo, Ailiena O. ^[2]; McBride, Molly J. ^[1]; Palowitch, Gavin M. ^[1]; Pandelia, Maria-Eirini ^[3]; ^[1]; Yennawar, Neela H. ^[1]; ^[1]

Pennsylvania State Univ., University Park, PA (United States)
Pennsylvania State Univ., University Park, PA (United States); California Institute of Technology (CalTech), Pasadena, CA (United States)
Brandeis Univ., Waltham, MA (United States)

Class I ribonucleotide reductases (RNRs) share a common mechanism of nucleotide reduction in a catalytic α subunit. All RNRs initiate catalysis with a thiyl radical, generated in class I enzymes by a metallocofactor in a separate β subunit. Class Id RNRs use a simple mechanism of cofactor activation involving oxidation of a Mn^II₂ cluster by free superoxide to yield a metal-based Mn^IIIMn^IV oxidant. This simple cofactor assembly pathway suggests that class Id RNRs may be representative of the evolutionary precursors to more complex class Ia–c enzymes. X-ray crystal structures of two class Id α proteins from Flavobacterium johnsoniae (Fj) and Actinobacillus ureae (Au) reveal that this subunit is distinctly small. The enzyme completely lacks common N-terminal ATP-cone allosteric motifs that regulate overall activity, a process that normally occurs by dATP-induced formation of inhibitory quaternary structures to prevent productive β subunit association. Class Id RNR activity is insensitive to dATP in the Fj and Au enzymes evaluated here, as expected. However, the class Id α protein from Fj adopts higher-order structures, detected crystallographically and in solution. The Au enzyme does not exhibit these quaternary forms. Our study reveals structural similarity between bacterial class Id and eukaryotic class Ia α subunits in conservation of an internal auxiliary domain. Overall, our findings with the Fj enzyme illustrate that nucleotide-independent higher-order quaternary structures can form in simple RNRs with truncated or missing allosteric motifs.

View Accepted Manuscript (DOE)

Research Organization:: Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)

Sponsoring Organization:: Michigan Economic Development Corporation; NIH High-End Instrumentation Grant; National Cancer Institute; National Institutes for Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1544859

Journal Information:: Biochemistry, Journal Name: Biochemistry Journal Issue: 14 Vol. 58; ISSN 0006-2960

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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