The manganese-containing ribonucleotide reductase of Corynebacterium ammoniagenes is a class Ib enzyme

Franck Fieschi, Eduard Torrents, Larisa Toulokhonova, Albert Jordan, Ulf Hellman, Jordi Barbe, Isidre Gibert, Margareta Karlsson, Britt Marie Sjöberg

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45 Citations (Scopus)

Abstract

Ribonucleotide reductases (RNRs) are key enzymes in living cells that provide the precursors of DNA synthesis. The three characterized classes of RNRs differ by their metal cofactor and their stable organic radical. We have purified to near homogeneity the enzymatically active Mn-containing RNR of Corynebacterium ammoniagenes, previously claimed to represent a fourth RNR class. N-terminal and internal peptide sequence analyses clearly indicate that the C. ammoniagenes RNR is a class lb enzyme. In parallel, we have cloned a 10-kilo-base pair fragment from C. ammoniagenes genomic DNA, using primers specific for the known class Ib RNR. The cloned class Ib locus contains the nrdHIEF genes typical for class lb RNR operon. The deduced amino acid sequences of the nrdE and nrdF genes matched the peptides from the active enzyme, demonstrating that C. ammoniagenes RNR is composed of R1E and R2F components typical of class Ib. We also show that the Mn-containing RNR has a specificity for the NrdH-redoxin and a response to allosteric effectors that are typical of class Ib RNRs. Electron paramagnetic resonance and atomic absorption analyses confirm the presence of Mn as a cofactor and show, for the first time, insignificant amounts of iron and cobalt found in the other classes of RNR. Our discovery that C. ammoniagenes RNR is a class Ib enzyme and possesses all the highly conserved amino acid side chains that are known to ligate two ferric ions in other class I RNRs evokes new, challenging questions about the control of the metal site specificity in RNR. The cloning of the entire NrdHIEF locus of C. ammoniagenes will facilitate further studies along these lines.
Original languageEnglish
Pages (from-to)4329-4337
JournalJournal of Biological Chemistry
Volume273
DOIs
Publication statusPublished - 20 Feb 1998

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    Fieschi, F., Torrents, E., Toulokhonova, L., Jordan, A., Hellman, U., Barbe, J., Gibert, I., Karlsson, M., & Sjöberg, B. M. (1998). The manganese-containing ribonucleotide reductase of Corynebacterium ammoniagenes is a class Ib enzyme. Journal of Biological Chemistry, 273, 4329-4337. https://doi.org/10.1074/jbc.273.8.4329