Ribonucleotide reductase (RNR) of Corynebacterium glutamicum ATCC 13032-genetic characterization of a second class IV enzyme

• Published in: Microbiology (Society for General Microbiology) 1999-07, Vol.145 (7), p.1595-1604
• Main Authors: Oehlmann, Wulf, Auling, Georg
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Blotting, Southern; Cloning, Molecular; Corynebacterium - enzymology; Corynebacterium - genetics; Corynebacterium - growth & development; Corynebacterium glutamicum; DNA, Bacterial - analysis; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetic Complementation Test; Genetics; Microbiology; Molecular Sequence Data; nrdE gene; nrdF gene; Phylogeny; Restriction Mapping; Ribonucleotide Reductases - chemistry; Ribonucleotide Reductases - genetics; Ribonucleotide Reductases - metabolism; Sequence Analysis, DNA
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/13500872-145-7-1595

Institut für Mikrobiologie, Universität Hannover, Schneiderberg 50, D-30167 Hannover, Germany Author for correspondence: Georg Auling. Fax: -49 511 7625287. e-mail: [e-mail] ABSTRACT Ribonucleotide reductases (RNRs) encoded by nrd (nucleotide reduction) genes are unique enzymes providing the DNA precursors in all living organisms and several viruses. The designation of four classes of RNRs reflects their use of diverse metallo-cofactors. Using oligonucleotide primers derived from conserved domains of the primary structure of known NrdA and NrdE proteins, an internal 938 bp fragment of the nrdE gene was amplified from genomic DNA of Corynebacterium glutamicum. With this PCR product a 4·36 kb fragment was identified and cloned containing the nrdHIE genes of C. glutamicum. A probe derived from nrdF2 of Mycobacterium tuberculosis allowed the cloning and sequencing of the nrdF gene located 3·1 kb further downstream, encoding the small subunit of the C. glutamicum RNR. Conjugative introduction of nrdE from C. glutamicum complemented thermosensitive mutants of Corynebacterium ammoniagenes which had a defective catalytic subunit of the Mn-RNR. The authors provide arguments for allocation of the C. glutamicum NrdEF proteins to class IV in the RNR classification scheme of Stubbe & van der Donk (1995) [ Chem Biol 2, 793-801]. Keywords: Corynebacterium glutamicum, nrdHIE genes, distal nrdF gene, heterologous genetic complementation of nrd ts mutants, Mn-RNR of C. ammoniagenes The GenBank accession numbers for the sequences reported in this paper are AF112535 for nrdHIE and AF112536 for nrdF of C. glutamicum ATCC 13032.