Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1971 Oct;108(1):145–153. doi: 10.1128/jb.108.1.145-153.1971

Metabolism of 4-N-Hydroxy-Cytidine in Escherichia coli

R B Trimble 1, Frank Maley 1
PMCID: PMC247043  PMID: 4941553

Abstract

4-N-hydroxy-cytidine was found to substitute for uridine as a pyrimidine supplement for the growth of Escherichia coli Bu−. Measurement of the incorporation of 4-N-hydroxy-cytidine-2-14C into ribonucleic acid and deoxyribonucleic acid revealed that this compound was converted to cytidine or uridine before utilization. Two pathways for metabolism were considered: (i) the reduction of 4-N-hydroxy-cytidine to cytidine followed by deamination, (ii) the direct hydrolysis of hydroxylamine from 4-N-hydroxy-cytidine to yield uridine. A threefold increase in cytidine (deoxycytidine) deaminase (EC 3.5.4.5) activity, when the cells were grown on 4-N-hydroxy-cytidine, suggested the involvement of this enzyme. More direct proof was obtained by purifying the deaminase 185-fold and finding that it released hydroxylamine from 4-N-hydroxy-cytidine at one-fiftieth the rate at which ammonia was removed from cytidine. This result is consistent with the slower rate of growth of the Bu− cells on 4-N-hydroxy-cytidine than cytidine and suggests that the second pathway is the major route for utilization of this compound.

Full text

PDF
153

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BROWN D. M., SCHELL P. NUCLEOTIDES. 48. THE REACTION OF HYDROXYLAMINE WITH CYTOSINE AND RELATED COMPOUNDS. J Chem Soc. 1965 Jan;47:208–215. [PubMed] [Google Scholar]
  2. COHEN S. S., LICHTENSTEIN J., BARNER H. D., GREEN M. Studies on the biosynthesis of bacterial and viral pyrimidines. IV. Utilization of pyrimidine bases and nucleosides by bacterial mutants. J Biol Chem. 1957 Oct;228(2):611–619. [PubMed] [Google Scholar]
  3. FREESE E., BAUTZ-FREESE E., BAUTZ E. Hydroxylamine as a mutagenic and inactivating agent. J Mol Biol. 1961 Apr;3:133–143. doi: 10.1016/s0022-2836(61)80040-5. [DOI] [PubMed] [Google Scholar]
  4. FREESE E., BAUTZ E., FREESE E. B. The chemical and mutagenic specificity of hydroxylamine. Proc Natl Acad Sci U S A. 1961 Jun 15;47:845–855. doi: 10.1073/pnas.47.6.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fishbein W. N. Determination of hydroxylamine with Nessler reagent. Anal Chim Acta. 1967 Apr;37(4):484–489. doi: 10.1016/s0003-2670(01)80711-x. [DOI] [PubMed] [Google Scholar]
  6. Gerwin B. I., Stein W. H., Moore S. On the specificity of streptococcal proteinase. J Biol Chem. 1966 Jul 25;241(14):3331–3339. [PubMed] [Google Scholar]
  7. HECHT L. I., POTTER V. R. Nucleic acid metabolism in regenerating rat liver. I. The rate of deoxyribonucleic acid synthesis in vivo. Cancer Res. 1956 Nov;16(10 Pt 1):988–993. [PubMed] [Google Scholar]
  8. Hanze A. R. Nucleic acids. IV. The catalytic reduction of pyrimidine nucleosides (human liver deaminase inhibitors). J Am Chem Soc. 1967 Dec 6;89(25):6720–6725. doi: 10.1021/ja01001a057. [DOI] [PubMed] [Google Scholar]
  9. Ipata P. L., Cercignani G., Balestreri E. Partial purification and properties of cytidine deaminase from baker's yeast. Biochemistry. 1970 Aug 18;9(17):3390–3395. doi: 10.1021/bi00819a015. [DOI] [PubMed] [Google Scholar]
  10. Ipata Pier Luigi, Falcone Giuseppe, Serra Maria Cristina. Some regulatory properties of cytidine deaminase and uridine phosphorylase of Bacillus cereus. FEBS Lett. 1970 Sep 18;10(1):67–70. doi: 10.1016/0014-5793(70)80417-3. [DOI] [PubMed] [Google Scholar]
  11. Karlström O., Larsson A. Significance of ribonucleotide reduction in the biosynthesis of deoxyribonucleotides in Escherichia coli. Eur J Biochem. 1967 Dec;3(2):164–170. doi: 10.1111/j.1432-1033.1967.tb19512.x. [DOI] [PubMed] [Google Scholar]
  12. Klein A., Eisenstadt A., Eisenstadt J., Lengyel P. Inhibition of peptide-chain initiation in Escherichia coli by hydroxylamine and effects on ribonucleic acid synthesis. Biochemistry. 1970 Nov 10;9(23):4542–4549. doi: 10.1021/bi00825a013. [DOI] [PubMed] [Google Scholar]
  13. LAZZARINI R. A., ATKINSON D. E. A triphosphopyridine nucleotide-specific nitrite reductase from Escherichia coli. J Biol Chem. 1961 Dec;236:3330–3335. [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Maley F. The incorporation of precursors into chick embryo DNA in the presence of 4 N-hydroxy-deoxycytidine and 4 N-hydroxy-5-methyl-deoxycytidine. Biochem Z. 1965 Aug 19;342(4):420–428. [PubMed] [Google Scholar]
  16. Nelson D. J., Carter C. E. Inhibition of the biosynthesis of thymidylic acid by 4-N-Hydroxy-2'-deoxycytidine in L5178Y leukemic cells. Mol Pharmacol. 1966 May;2(3):248–258. [PubMed] [Google Scholar]
  17. Nixon P. F., Bertino J. R. Inhibition of peptide chain initiation in Escherichia coli by hydroxylamine. Reaction of hydroxylamine with folate coenzymes. Biochemistry. 1970 Dec 8;9(25):4833–4838. doi: 10.1021/bi00827a001. [DOI] [PubMed] [Google Scholar]
  18. O'Donovan G. A., Edlin G., Fuchs J. A., Neuhard J., Thomassen E. Deoxycytidine triphosphate deaminase: characterization of an Escherichia coli mutant deficient in the enzyme. J Bacteriol. 1971 Feb;105(2):666–672. doi: 10.1128/jb.105.2.666-672.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Donovan G. A., Neuhard J. Pyrimidine metabolism in microorganisms. Bacteriol Rev. 1970 Sep;34(3):278–343. doi: 10.1128/br.34.3.278-343.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ressler N. A simple and sensitive method for the measurement of deoxycytidylate deaminase activity. Clin Chem. 1969 Jul;15(7):575–581. [PubMed] [Google Scholar]
  21. WYATT G. R. The purine and pyrimidine composition of deoxypentose nucleic acids. Biochem J. 1951 May;48(5):584–590. doi: 10.1042/bj0480584. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES