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J. Biol. Chem., Vol. 276, Issue 2, 884-894, January 12, 2001
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From the Pyrimidine-requiring cdd mutants of
Escherichia coli deficient in cytidine deaminase utilize
cytidine as a pyrimidine source by an alternative pathway. This has
been presumed to involve phosphorylation of cytidine to CMP by
cytidine/uridine kinase and subsequent hydrolysis of CMP to cytosine
and ribose 5-phosphate by a putative CMP hydrolase. Here we show that
cytidine, in cdd strains, is converted directly to cytosine
and ribose by a ribonucleoside hydrolase encoded by the previously
uncharacterized gene ybeK, which we have renamed rihA. The RihA enzyme is homologous to the products of two
unlinked genes, yeiK and yaaF, which have been
renamed rihB and rihC, respectively. The RihB
enzyme was shown to be a pyrimidine-specific ribonucleoside hydrolase
like RihA, whereas RihC hydrolyzed both pyrimidine and purine
ribonucleosides. The physiological function of the ribonucleoside hydrolases in wild-type E. coli strains is enigmatic, as
their activities are paralleled by the phosphorolytic activities of the
nucleoside phosphorylases, and a triple mutant lacking all three
hydrolytic activities grew normally. Furthermore, enzyme assays and
lacZ gene fusion analysis indicated that rihB
was essentially silent unless activated by mutation, whereas
rihA and rihC were poorly expressed in glucose
medium due to catabolite repression.
The RihA, RihB, and RihC Ribonucleoside Hydrolases of
Escherichia coli
SUBSTRATE SPECIFICITY, GENE EXPRESSION, AND REGULATION*
§ and Department of Biological Chemistry,
Institute of Molecular Biology, University of Copenhagen, Sølvgade
83H, DK1307 Copenhagen K, Denmark and the ¶ John F. Kennedy
Institute, Gammel Landevej 7, 2600 Glostrup, Denmark
*
This work was supported by the Danish Natural Science
Research Council, the Danish Health Insurance Foundation, and the
Foundation of 1870.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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