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. 1969 Jul;63(3):904–910. doi: 10.1073/pnas.63.3.904

REPLICATION AND PROPERTIES OF DNA IN NICOTINAMIDE ADENINE DINUCLEOTIDE DEFICIENCY OF Escherichia coli CELLS

Ryushi Nozawa 1, Den'ichi Mizuno 1
PMCID: PMC223538  PMID: 4310516

Abstract

When niacin-requiring E. coli was starved of niacin, a precursor of nicotinamide adenine dinucleotide (NAD), the cells elongated and synthesis of DNA was inhibited while syntheses of RNA and protein continued. The cellular NAD concentration decreased to less than one tenth of the normal level in niacin deficiency, but no change in DNA ligase level was detected. The apparent inhibition of DNA synthesis can be explained as due to reduction in the activity of the cofactor, NAD, for the enzyme.

The DNA synthesized in this state was small in an alkaline sucrose gradient but of normal size in a neutral sucrose gradient. The molecules had a double-stranded structure, as judged by hydroxyapatite chromatography. The small pieces of DNA were integrated into normal-size DNA molecules on addition of niacin, when the cellular NAD concentration was restored to the normal level but DNA synthesis was not. It is suggested that this small DNA is an intermediate in DNA replication.

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Selected References

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

  1. Becker A., Lyn G., Gefter M., Hurwitz J. The enzymatic repair of DNA, II. Characterization of phage-induced sealase. Proc Natl Acad Sci U S A. 1967 Nov;58(5):1996–2003. doi: 10.1073/pnas.58.5.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cozzarelli N. R., Melechen N. E., Jovin T. M., Kornberg A. Polynucleotide cellulose as a substrate for a polynucleotide ligase induced by phage T4. Biochem Biophys Res Commun. 1967 Aug 23;28(4):578–586. doi: 10.1016/0006-291x(67)90353-1. [DOI] [PubMed] [Google Scholar]
  3. Gefter M. L., Becker A., Hurwitz J. The enzymatic repair of DNA. I. Formation of circular lambda-DNA. Proc Natl Acad Sci U S A. 1967 Jul;58(1):240–247. doi: 10.1073/pnas.58.1.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Helmstetter C. E., Pierucci O. Cell division during inhibition of deoxyribonucleic acid synthesis in Escherichia coli. J Bacteriol. 1968 May;95(5):1627–1633. doi: 10.1128/jb.95.5.1627-1633.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hosoda J., Mathews E. DNA replication in vivo by a temperature-sensitive polynucleotide ligase mutant of T4. Proc Natl Acad Sci U S A. 1968 Nov;61(3):997–1004. doi: 10.1073/pnas.61.3.997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Iizuka M., Mizuno D. Turnover of NAD in bacteria. Biochim Biophys Acta. 1967 Nov 28;148(2):320–327. doi: 10.1016/0304-4165(67)90126-2. [DOI] [PubMed] [Google Scholar]
  7. Lindahl T., Edelman G. M. Polynucleotide ligase from myeloid and lymphoid tissues. Proc Natl Acad Sci U S A. 1968 Oct;61(2):680–687. doi: 10.1073/pnas.61.2.680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. London J., Knight M. Concentrations of nicotinamide nucleotide coenzymes in micro-organisms. J Gen Microbiol. 1966 Aug;44(2):241–254. doi: 10.1099/00221287-44-2-241. [DOI] [PubMed] [Google Scholar]
  9. MIYAZAWA Y., THOMAS C. A., Jr NUCLEOTIDE COMPOSITION OF SHORT SEGMENTS OF DNA MOLECULES. J Mol Biol. 1965 Feb;11:223–237. doi: 10.1016/s0022-2836(65)80053-5. [DOI] [PubMed] [Google Scholar]
  10. Newman J., Hanawalt P. Role of polynucleotide ligase in T4 DNA replication. J Mol Biol. 1968 Aug 14;35(3):639–642. doi: 10.1016/s0022-2836(68)80020-8. [DOI] [PubMed] [Google Scholar]
  11. Nozawa R., Natori S., Obinata M., Mizuno D. DNA synthesis during niacin starvation in niacin requiring Escherichia coli. Biochem Biophys Res Commun. 1968 Aug 21;32(4):602–609. doi: 10.1016/0006-291x(68)90280-5. [DOI] [PubMed] [Google Scholar]
  12. Oishi M. Studies of DNA replication in vivo. I. Isolation of the first intermediate of DNA replication in bacteria as single-stranded DNA. Proc Natl Acad Sci U S A. 1968 May;60(1):329–336. doi: 10.1073/pnas.60.1.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Okazaki R., Okazaki T., Sakabe K., Sugimoto K., Sugino A. Mechanism of DNA chain growth. I. Possible discontinuity and unusual secondary structure of newly synthesized chains. Proc Natl Acad Sci U S A. 1968 Feb;59(2):598–605. doi: 10.1073/pnas.59.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Olivera B. M., Lehman I. R. Diphosphopyridine nucleotide: a cofactor for the polynucleotide-joining enzyme from Escherichia coli. Proc Natl Acad Sci U S A. 1967 Jun;57(6):1700–1704. doi: 10.1073/pnas.57.6.1700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Olivera B. M., Lehman I. R. Linkage of polynucleotides through phosphodiester bonds by an enzyme from Escherichia coli. Proc Natl Acad Sci U S A. 1967 May;57(5):1426–1433. doi: 10.1073/pnas.57.5.1426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pauling C., Hamm L. Properties of a temperature-sensitive radiation-sensitive mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1495–1502. doi: 10.1073/pnas.60.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Richardson C. C. Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli. Proc Natl Acad Sci U S A. 1965 Jul;54(1):158–165. doi: 10.1073/pnas.54.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rupp W. D., Howard-Flanders P. Discontinuities in the DNA synthesized in an excision-defective strain of Escherichia coli following ultraviolet irradiation. J Mol Biol. 1968 Jan 28;31(2):291–304. doi: 10.1016/0022-2836(68)90445-2. [DOI] [PubMed] [Google Scholar]
  19. Sugimoto K., Okazaki T., Okazaki R. Mechanism of DNA chain growth, II. Accumulation of newly synthesized short chains in E. coli infected with ligase-defective T4 phages. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1356–1362. doi: 10.1073/pnas.60.4.1356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weiss B., Richardson C. C. Enzymatic breakage and joining of deoxyribonucleic acid, I. Repair of single-strand breaks in DNA by an enzyme system from Escherichia coli infected with T4 bacteriophage. Proc Natl Acad Sci U S A. 1967 Apr;57(4):1021–1028. doi: 10.1073/pnas.57.4.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yudelevich A., Ginsberg B., Hurwitz J. Discontinuous synthesis of DNA during replication. Proc Natl Acad Sci U S A. 1968 Nov;61(3):1129–1136. doi: 10.1073/pnas.61.3.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zimmerman S. B., Little J. W., Oshinsky C. K., Gellert M. Enzymatic joining of DNA strands: a novel reaction of diphosphopyridine nucleotide. Proc Natl Acad Sci U S A. 1967 Jun;57(6):1841–1848. doi: 10.1073/pnas.57.6.1841. [DOI] [PMC free article] [PubMed] [Google Scholar]

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