1887

Abstract

Genes encoding two ribonucleotide reductases (RNRs) were identified in members of the genus . One gene, , encoded an oligomeric protein comprising four identical subunits each with a molecular mass of ∼108 kDa. The activity of this protein depended on the presence of 5′-deoxyadenosylcobalamine (coenzyme B), establishing it as a class II RNR. The gene was cloned, using internal peptide sequences from the purified protein, and was found to encode a polypeptide of 961 aa. Molecular phylogenetic analysis showed that the class II RNR shares significant similarity with most other bacterial and archaeal class II RNRs. Two other genes, and , were initially identified in the genome database in unannotated ORFs as encoding a class Ia RNR. Southern analysis demonstrated that the genes were present in different spp. The genes were cloned and expressed in , and the recombinant proteins were shown to represent a class I RNR. It was shown, using quantitative real-time PCR, that the class Ia and class II RNR genes were differentially transcribed during vegetative growth. The copy number of the class II transcripts was approximately constant throughout the exponential phase of vegetative growth (3–5×10 copies per 400 ng total RNA after reverse transcription). In contrast, the copy number of the class Ia transcripts was some 10- to 20-fold less than that of in the early-exponential growth phase (28×10 copies), and decreased markedly at the mid-exponential (4×10 copies) and late-exponential phases (11×10 copies) of growth. A possible role for the involvement of two RNRs during vegetative growth is discussed.

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2002-02-01
2020-03-29
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