Discovery and characterization of a novel uridine hydrolase

Abstract

Modified nucleotides are present in various RNA species. They have great chemical diversity and are critical for RNA folding, stability, and interaction with cellular proteins. Biosynthesis of pyrimidine nucleotides and their modified derivatives in RNA is well studied. Still, much less is known about the cellular degradation of these compounds and their salvage. Using bacterial genetics methods, an E. coli strain, unable to convert 2’-O-methyluridine into uracil and therefore to grow on minimal medium, was created. The growth phenotype was restored by introducing a genome fragment from the metagenomic libraries to this bacterium. This fragment contains 2 ORFs, one encoding a bacterial nucleotide hydrolase, and the other – a ribose aldolase. The recombinant nucleotide hydrolase was expressed in E. coli, purified, and its substrate specificity was demonstrated. This work is supported by the Research Council of Lithuania (LMT, SEN-07/2015).