Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Apr;150(1):286–292. doi: 10.1128/jb.150.1.286-292.1982

dnaC-dependent reconstitution of replication forks in Escherichia coli lysates.

V Nüsslein-Crystalla, I Niedenhof, R Rein
PMCID: PMC220111  PMID: 6277866

Abstract

Lysates of Escherichia coli exhibit a DNA-synthesizing activity that depends on the presence of replication forks and of replication proteins. Replicative activity was reconstituted in vitro by mixing lysates prepared from temperature-sensitive dnaB mutants with wild-type dnaB protein. Lysates of double mutants deficient in both dnaB and dnaC genes could only be complemented by the addition of both dnaB and dnaC proteins, whereas lysates deficient in dnaC protein did not require the addition of any exogenous factor. This shows that the replication machinery, once it is running along the chromosome, is independent of dnaC protein, dnaC activity, however, is required for the replacement of defective dnaB protein at running replication forks.

Full text

PDF
286

Images in this article

289Image
on p.289

Selected References

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

  1. Alberts B., Sternglanz R. Recent excitement in the DNA replication problem. Nature. 1977 Oct 20;269(5630):655–661. doi: 10.1038/269655a0. [DOI] [PubMed] [Google Scholar]
  2. Carl P. L. Escherichia coli mutants with temperature-sensitive synthesis of DNA. Mol Gen Genet. 1970;109(2):107–122. doi: 10.1007/BF00269647. [DOI] [PubMed] [Google Scholar]
  3. Glassberg J., Meyer R. R., Kornberg A. Mutant single-strand binding protein of Escherichia coli: genetic and physiological characterization. J Bacteriol. 1979 Oct;140(1):14–19. doi: 10.1128/jb.140.1.14-19.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hübscher U., Kornberg A. The delta subunit of Escherichia coli DNA polymerase III holoenzyme is the dnaX gene product. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6284–6288. doi: 10.1073/pnas.76.12.6284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kung F. C., Glaser D. A. dnaA acts before dnaC in the initiation of DNA replication. J Bacteriol. 1978 Feb;133(2):755–762. doi: 10.1128/jb.133.2.755-762.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Low R. L., Arai K., Kornberg A. Conservation of the primosome in successive stages of phi X174 DNA replication. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1436–1440. doi: 10.1073/pnas.78.3.1436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. MAALOE O., HANAWALT P. C. Thymine deficiency and the normal DNA replication cycle. I. J Mol Biol. 1961 Apr;3:144–155. doi: 10.1016/s0022-2836(61)80041-7. [DOI] [PubMed] [Google Scholar]
  8. Marsh R. C., Worcel A. A DNA fragment containing the origin of replication of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2720–2724. doi: 10.1073/pnas.74.7.2720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Messer W. Initiation of deoxyribonucleic acid replication in Escherichia coli B-r: chronology of events and transcriptional control of initiation. J Bacteriol. 1972 Oct;112(1):7–12. doi: 10.1128/jb.112.1.7-12.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nüsslein-Crystalla V., Scheefers-Borchel U. In vitro replication of a DNA fragment containing the vicinity of the origin of E. coli DNA replication. Mol Gen Genet. 1979 Jan 16;169(1):35–40. doi: 10.1007/BF00267542. [DOI] [PubMed] [Google Scholar]
  11. Nüsslein V., Henke S., Johnston L. H. Replication of E. coli duplex DNA in vitro. The separation of the DNA containing fractions of a lysate from the soluble enzymes and their complementation properties. Mol Gen Genet. 1976 May 7;145(2):183–190. doi: 10.1007/BF00269592. [DOI] [PubMed] [Google Scholar]
  12. Schaller H., Otto B., Nüsslein V., Huf J., Herrmann R., Bonhoeffer F. Deoxyribonucleic acid replication in vitro. J Mol Biol. 1972 Jan 28;63(2):183–200. doi: 10.1016/0022-2836(72)90369-5. [DOI] [PubMed] [Google Scholar]
  13. Schuster H., Schlicht M., Lanka E., Mikolajczyk M., Edelbluth C. DNA synthesis in an Escherichia coli dna B dnaC mutant. Mol Gen Genet. 1977 Feb 28;151(1):11–16. doi: 10.1007/BF00446907. [DOI] [PubMed] [Google Scholar]
  14. Sevastopoulos C. G., Wehr C. T., Glaser D. A. Large-scale automated isolation of Escherichia coli mutants with thermosensitive DNA replication. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3485–3489. doi: 10.1073/pnas.74.8.3485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Staudenbauer W. L., Lanka E., Schuster H. Replication of small plasmids in extracts of Escherichia coli: involvement of the dnaB and dnaC protein in the replication of early replicative intermediates. Mol Gen Genet. 1978 Jul 4;162(3):243–249. doi: 10.1007/BF00268849. [DOI] [PubMed] [Google Scholar]
  16. Sueoka N., Yoshikawa H. The chromosome of Bacillus subtilis. I. Theory of marker frequency analysis. Genetics. 1965 Oct;52(4):747–757. doi: 10.1093/genetics/52.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wechsler J. A. Genetic and phenotypic characterization of dnaC mutations. J Bacteriol. 1975 Feb;121(2):594–599. doi: 10.1128/jb.121.2.594-599.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]
  19. Wechsler J. A., Nüsslein V., Otto B., Klein A., Bonhoeffer F., Herrmann R., Gloger L., Schaller H. Isolation and characterization of thermosensitive Escherichia coli mutants defective in deoxyribonucleic acid replication. J Bacteriol. 1973 Mar;113(3):1381–1388. doi: 10.1128/jb.113.3.1381-1388.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wickner S. H. DNA replication proteins of Escherichia coli. Annu Rev Biochem. 1978;47:1163–1191. doi: 10.1146/annurev.bi.47.070178.005503. [DOI] [PubMed] [Google Scholar]
  21. Wickner S., Berkower I., Wright M., Hurwitz J. Studies on in vitro DNA synthesis: purification of dna C gene product containing dna D activity from Escherichia coli. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2369–2373. doi: 10.1073/pnas.70.8.2369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zyskind J. W., Deen L. T., Smith D. W. Temporal sequence of events during the initiation process in Escherichia coli deoxyribonucleic acid replication: roles of the dnaA and dnaC gene products and ribonucleic acid polymerase. J Bacteriol. 1977 Mar;129(3):1466–1475. doi: 10.1128/jb.129.3.1466-1475.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES