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. 1979 Jan;29(1):61–68. doi: 10.1128/jvi.29.1.61-68.1979

Deoxythymidine nucleotide metabolism in Bacillus subtilis W23 infected with bacteriophage SP1Oc: preliminary evidence that dTMP in SP10c DNA is synthesized by a novel, bacteriophage-specific mechanism.

O Markewych, E Casella, M Dosmar, H Witmer
PMCID: PMC353073  PMID: 107324

Abstract

Despite the fact that mature SP10c DNA contains dTMP, the acid-soluble fraction of infected cells contained no dTTP during the interval of phage replication. However, infected cells contained normal cellular levels of dATP, dGTP, and dCTP. Upon infection of deoxythymidine-starved Bacillus subtilis M160 (a deoxythymidine-requiring mutant of B. subtilis W23), mature phage DNA with a normal dTMP content was made. SP10c codes for an enzyme that seems to catalyze the tetrahydrofolate-dependent transfer of 1-carbon fragments to the 5 position of dUMP. The transfer of 1-carbon fragments is not accompanied by oxidation of tetrahydrofolage to dihydrofolate, implying that the enzyme in question is not a dTMP synthetase. It is proposed that dTMP in mature SP10c DNA is derived by the postreplicational modification of some other nucleotide and not by the direct incorporation of dTTP into DNA.

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Selected References

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

  1. Ando T. A nuclease specific for heat-denatured DNA in isolated from a product of Aspergillus oryzae. Biochim Biophys Acta. 1966 Jan 18;114(1):158–168. doi: 10.1016/0005-2787(66)90263-2. [DOI] [PubMed] [Google Scholar]
  2. BOTT K., STRAUSS B. THE CARRIER STATE OF BACILLUS SUBTILIS INFECTED WITH THE TRANSDUCING BACTERIOPHAGE SP10. Virology. 1965 Feb;25:212–225. doi: 10.1016/0042-6822(65)90200-x. [DOI] [PubMed] [Google Scholar]
  3. Baros A., Witmer H. J. Effect of chloramphenicol and starvation for an essential amino acid on the synthesis and decay of T4 bacteriophage-specific messengers transcribed from early and quasi-late promoters. Arch Biochem Biophys. 1975 Aug;169(2):415–427. doi: 10.1016/0003-9861(75)90183-6. [DOI] [PubMed] [Google Scholar]
  4. Brandon C., Gallop P. M., Marmur J., Hayashi H., Nakanishi K. Structure of a new pyrimidine from Bacillus subtilis phage SP-15 nucleic acid. Nat New Biol. 1972 Sep 20;239(90):70–71. doi: 10.1038/newbio239070a0. [DOI] [PubMed] [Google Scholar]
  5. Brown N. C. 6-(p-hydroxyphenylazo)-uracil: a selective inhibitor of host DNA replication in phage-infected Bacillus subtilis. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1454–1461. doi: 10.1073/pnas.67.3.1454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Budman D. R., Pardee A. B. Thymidine and thymine incorporation into deoxyribonucleic acid: inhibition and repression by uridine of thymidine phosphorylase of Escherichia coli. J Bacteriol. 1967 Nov;94(5):1546–1550. doi: 10.1128/jb.94.5.1546-1550.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hemphill H. E., Whiteley H. R. Bacteriophages of Bacillus subtilis. Bacteriol Rev. 1975 Sep;39(3):257–315. doi: 10.1128/br.39.3.257-315.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kelln R. A., Warren R. A. Studies on the biosynthesis of alpha-putrescinylthymine in bacteriophage phi W-14-infected Pseudomonas acidovorans. J Virol. 1973 Dec;12(6):1427–1433. doi: 10.1128/jvi.12.6.1427-1433.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kropinski A. M., Bose R. J., Warren R. A. 5-(4-Aminobutylaminomethyl)uracil, an unusual pyrimidine from the deoxyribonucleic acid of bacteriophage phiW-14. Biochemistry. 1973 Jan 2;12(1):151–157. doi: 10.1021/bi00725a025. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Lomax M. I., Greenberg G. R. A new assay of thymidylate synthetase activity based on the release of tritium from deoxyuridylate-5-3-H. J Biol Chem. 1967 Jan 10;242(1):109–113. [PubMed] [Google Scholar]
  12. Markewych O., Boghosian A., Dosmar M., Ende D., Witmer H. SP-10 bacteriophage-specific nucleic acid and enzyme synthesis in Bacillus subtilis W23. J Virol. 1977 Jan;21(1):84–95. doi: 10.1128/jvi.21.1.84-95.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Neubort S., Marmur J. Synthesis of the unusual DNA of Bacillus subtilis bacteriophage SP-15. J Virol. 1973 Nov;12(5):1078–1084. doi: 10.1128/jvi.12.5.1078-1084.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Neuhard J., Price A. R., Schack L., Thomassen E. Two thymidylate synthetases in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1194–1198. doi: 10.1073/pnas.75.3.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Neville M. M., Brown N. C. Inhibition of a discrete bacterial DNA polymerase by 6-(p-hydroxyphenylazo)-uracil and 6-(p-hydroxyphenylazo-)-isocytosine. Nat New Biol. 1972 Nov 15;240(98):80–82. doi: 10.1038/newbio240080a0. [DOI] [PubMed] [Google Scholar]
  16. Oka J., Ueda K., Hayaishi O. Snake venom phosphodiesterase: simple purification with Blue Sepharose and its application to poly(ADP-ribose) study. Biochem Biophys Res Commun. 1978 Feb 28;80(4):841–848. doi: 10.1016/0006-291x(78)91321-9. [DOI] [PubMed] [Google Scholar]
  17. Plagemann P. G. Nucleotide pools of Novikoff rat hepatoma cells growing in suspension culture. I. Kinetics of incorporation of nucleosides into nucleotide pools and pool sizes during growth cycle. J Cell Physiol. 1971 Apr;77(2):213–240. doi: 10.1002/jcp.1040770212. [DOI] [PubMed] [Google Scholar]
  18. Sutton W. D. A crude nuclease preparation suitable for use in DNA reassociation experiments. Biochim Biophys Acta. 1971 Jul 29;240(4):522–531. doi: 10.1016/0005-2787(71)90709-x. [DOI] [PubMed] [Google Scholar]
  19. THORNE C. B. Transduction in Bacillus subtilis. J Bacteriol. 1962 Jan;83:106–111. doi: 10.1128/jb.83.1.106-111.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. WAHBA A. J., FRIEDKIN M. The enzymatic synthesis of thymidylate. I. Early steps in the purification of thymidylate synthetase of Escherichia coli. J Biol Chem. 1962 Dec;237:3794–3801. [PubMed] [Google Scholar]
  21. WIBERG J. S., DIRKSEN M. L., EPSTEIN R. H., LURIA S. E., BUCHANAN J. M. Early enzyme synthesis and its control in E. coli infected with some amber mutants of bacteriophage T4. Proc Natl Acad Sci U S A. 1962 Feb;48:293–302. doi: 10.1073/pnas.48.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Walker M. S., Mandel M. Biosynthesis of 5-(4'5'-dihydroxypentyl) uracil as a nucleoside triphosphate in bacteriophage SP15-infected Bacillus subtilis. J Virol. 1978 Feb;25(2):500–509. doi: 10.1128/jvi.25.2.500-509.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wiberg J. S., Mendelsohn S., Warner V., Hercules K., Aldrich C., Munro J. L. SP62, a viable mutant of bacteriophage T4D defective in regulation of phage enzyme synthesis. J Virol. 1973 Oct;12(4):775–792. doi: 10.1128/jvi.12.4.775-792.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Witmer H., Baros A., Forbes J., Padnos D., Maricondia W., Weiner M. Transcriptional control of T4 coliphage-specific genes 30, 42, 43, rIIA, rIIB, and e. J Gen Virol. 1976 Jun;31(3):289–302. doi: 10.1099/0022-1317-31-3-289. [DOI] [PubMed] [Google Scholar]
  25. Yagil E., Rosner A. Phosphorolysis of 5-fluoro-2'-deoxyuridine in Escherichia coli and its inhibition by nucleosides. J Bacteriol. 1971 Nov;108(2):760–764. doi: 10.1128/jb.108.2.760-764.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]

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