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. 1977 Mar;129(3):1476–1486. doi: 10.1128/jb.129.3.1476-1486.1977

NOVEL Escherichia coli dnaB mutant: direct involvement of the dnaB252 gene product in the synthesis of an origin-ribonucleic acid species during initiaion of a round of deoxyribonucleic acid replication.

J W Zyskind, D W Smith
PMCID: PMC235125  PMID: 321430

Abstract

The initiation process of deoxyribonucleic acid (DNA) replication in Escherichia coli has been studied using the thermoreversible dna initiation mutant E. coli HfrHl65/120/6 dna-252. This dna mutation was incorrectly classed as a dnaA mutation. Biochemical and genetic evidence suggests that the dna-252 mutant is a novel dnaB mutant, possessing phenotypic properties which distinguish it from other dnaB mutants. Sensitivity of reinitiation in the dna-252 mutant to specific inhibitors of protein, ribonucleic acid (RNA), and DNA synthesis was studied. Reinitiation is shown to be sensitive to rifampin and streptolydigin but not to cholramphenicol. Thus, the dna-252 gene product appears to be required during the initiation process for a step occurring either before or during synthesis of an RNA species (origin-RNA). Using reversible inhibition of RNA synthesis by streptolydigin of a streptolydigin-sensitive derivative of the dna-252 mutant, the dna-252 gene product is shown to be directly involved in the synthesis of an orgin-RNA species. These results are included in a schematic model presented in the accompanying paper of the temporal sequence of events occurring during the initiation process.

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Selected References

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  1. BONHOEFFER F., SCHALLER H. A METHOD FOR SELECTIVE ENRICHMENT OF MUTANTS BASED ON THE HIGH UV SENSITIVITY OF DNA CONTAINING 5-BROMOURACIL. Biochem Biophys Res Commun. 1965 Jun 18;20:93–97. [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beyersmann D., Schlicht M., Schuster H. Temperature-sensitive initiation of DNA replication in a mutant of Escherichia coli K12. Mol Gen Genet. 1971;111(2):145–158. doi: 10.1007/BF00267789. [DOI] [PubMed] [Google Scholar]
  4. Beyersmann D., Schuster H. DNA synthesis in P1 infected E. coli mutants temperature-sensitive in DNA replication. Mol Gen Genet. 1972;114(2):173–176. doi: 10.1007/BF00332788. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Cassani G., Burgess R. R., Goodman H. M., Gold L. Inhibition of RNA polymerase by streptolydigin. Nat New Biol. 1971 Apr 14;230(15):197–200. doi: 10.1038/newbio230197a0. [DOI] [PubMed] [Google Scholar]
  7. Clarke L., Carbon J. Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4361–4365. doi: 10.1073/pnas.72.11.4361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clewell D. B., Evenchik B. G. Effects of rifampicin, streptolydigin and actinomycin D on the replication of Col E1 plasmid DNA in Escherichia coli. J Mol Biol. 1973 Apr 15;75(3):503–513. doi: 10.1016/0022-2836(73)90457-9. [DOI] [PubMed] [Google Scholar]
  9. Collins J., Williams P., Helinski D. R. Plasmid ColE1 DNA replication in Escherichia coli strains temperature-sensitive for DNA replication. Mol Gen Genet. 1975;136(4):273–289. doi: 10.1007/BF00341713. [DOI] [PubMed] [Google Scholar]
  10. Eisenberg S., Scott J. F., Kornberg A. Enzymatic replication of viral and complementary strands of duplex DNA of phage phiX174 proceeds by seprate mechanisms. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3151–3155. doi: 10.1073/pnas.73.9.3151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fangman W. L., Novick A. Characterization of two bacterial mutants with temperature-sensitive synthesis of DNA. Genetics. 1968 Sep;60(1):1–17. doi: 10.1093/genetics/60.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goebel W. Studies on extrachromosomal DNA elements. Replication of the colicinogenic factor Col E1 in two temperature sensitive mutants of Escherichia coli defective in DNA replication. Eur J Biochem. 1970 Aug;15(2):311–320. doi: 10.1111/j.1432-1033.1970.tb01009.x. [DOI] [PubMed] [Google Scholar]
  13. Gross J. D. DNA replication in bacteria. Curr Top Microbiol Immunol. 1972;57:39–74. doi: 10.1007/978-3-642-65297-4_2. [DOI] [PubMed] [Google Scholar]
  14. Hanna M. H., Carl P. L. Reinitiation of deoxyribonucleic acid synthesis by deoxyribonucleic acid initiation mutants of Escherichia coli: role of ribonucleic acid synthesis, protein synthesis, and cell division. J Bacteriol. 1975 Jan;121(1):219–226. doi: 10.1128/jb.121.1.219-226.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Haselkorn R., Vogel M., Brown R. D. Conservation of the rifamycin sensitivity of transcription during T4 development. Nature. 1969 Mar 1;221(5183):836–838. doi: 10.1038/221836a0. [DOI] [PubMed] [Google Scholar]
  16. Kogoma T., Lark K. G. Characterization of the replication of Escherichia coli DNA in the absence of protein synthesis: stable DNA replication. J Mol Biol. 1975 May 15;94(2):243–256. doi: 10.1016/0022-2836(75)90081-9. [DOI] [PubMed] [Google Scholar]
  17. Kogoma T. Two types of temperature sensitivity in DNA replication of an Escherichia coli dnaB mutant. J Mol Biol. 1976 May 5;103(1):191–197. doi: 10.1016/0022-2836(76)90059-0. [DOI] [PubMed] [Google Scholar]
  18. Lanka E., Schuster H. Replication of bacteriophages in Escherichia coli mutants thermosensitive in DNA synthesis. Mol Gen Genet. 1970;106(3):274–285. doi: 10.1007/BF00340386. [DOI] [PubMed] [Google Scholar]
  19. Lark K. G. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. doi: 10.1016/0022-2836(72)90320-8. [DOI] [PubMed] [Google Scholar]
  20. Lark K. G., Wechsler J. A. DNA replication in dnaB mutants of Escherichia coli: gene product interaction and synthesis of 4 S pieces. J Mol Biol. 1975 Feb 15;92(1):145–163. doi: 10.1016/0022-2836(75)90095-9. [DOI] [PubMed] [Google Scholar]
  21. Laurent S. J. Initiation of deoxyribonucleic acid replication in a temperature-sensitive mutant of B. subtilis: evidence for a transcriptional step. J Bacteriol. 1973 Oct;116(1):141–145. doi: 10.1128/jb.116.1.141-145.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Low B. Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Reid P., Speyer J. Rifampicin inhibition of ribonucleic acid and protein synthesis in normal and ethylenediaminetetraacetic acid-treated Escherichia coli. J Bacteriol. 1970 Oct;104(1):376–389. doi: 10.1128/jb.104.1.376-389.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ryder O. A., Smith D. W. Isolation of membrane-associated folded chromosomes from Escherichia coli: effect of protein synthesis inhibition. J Bacteriol. 1974 Dec;120(3):1356–1363. doi: 10.1128/jb.120.3.1356-1363.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schekman R., Weiner A., Kornberg A. Multienzyme systems of DNA replication. Science. 1974 Dec 13;186(4168):987–993. doi: 10.1126/science.186.4168.987. [DOI] [PubMed] [Google Scholar]
  28. Schleif R. Isolation and characterization of streptolydigin resistant RNA polymerase. Nature. 1969 Sep 6;223(5210):1068–1069. doi: 10.1038/2231068a0. [DOI] [PubMed] [Google Scholar]
  29. Schuster H., Beyersmann D., Mikolajczyk M., Schlicht M. Prophage induction by high temperature in thermosensitive dna mutants lysogenic for bacteriophage lambda. J Virol. 1973 Jun;11(6):879–885. doi: 10.1128/jvi.11.6.879-885.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. 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]
  33. Wehrli W., Knüsel F., Schmid K., Staehelin M. Interaction of rifamycin with bacterial RNA polymerase. Proc Natl Acad Sci U S A. 1968 Oct;61(2):667–673. doi: 10.1073/pnas.61.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wehrli W., Staehelin M. Actions of the rifamycins. Bacteriol Rev. 1971 Sep;35(3):290–309. doi: 10.1128/br.35.3.290-309.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Weiner J. H., McMacken R., Kornberg A. Isolation of an intermediate which precedes dnaG RNA polymerase participation in enzymatic replication of bacteriophage phi X174 DNA. Proc Natl Acad Sci U S A. 1976 Mar;73(3):752–756. doi: 10.1073/pnas.73.3.752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wickner R. B., Wright M., Wickner S., Hurwitz J. Conversion of phiX174 and fd single-stranded DNA to replicative forms in extracts of Escherichia coli. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3233–3237. doi: 10.1073/pnas.69.11.3233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wickner S., Hurwitz J. Interaction of Escherichia coli dnaB and dnaC(D) gene products in vitro. Proc Natl Acad Sci U S A. 1975 Mar;72(3):921–925. doi: 10.1073/pnas.72.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wickner S., Wright M., Hurwitz J. Association of DNA-dependent and -independent ribonucleoside triphosphatase activities with dnaB gene product of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Mar;71(3):783–787. doi: 10.1073/pnas.71.3.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]

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