Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1973 Aug;70(8):2369–2373. doi: 10.1073/pnas.70.8.2369

Studies on In Vitro DNA Synthesis Purification of dna C Gene Product Containing dna D Activity from Escherichia coli*

Sue Wickner 1, Ira Berkower 1, Michel Wright 1, Jerard Hurwitz 1
PMCID: PMC433737  PMID: 4599623

Abstract

The conversion of ϕX174 single-stranded DNA to duplex DNA by extracts of E. coli requires products of the E. coli DNA replication genes. By use of this complementation system, the dna C gene product has been purified from wild-type E. coli as well as from a dna C temperature-sensitive mutant. The latter preparations are temperature sensitive when compared to the wild-type gene product. The dna C and dna D gene products copurify, have similar characteristics, are both temperature sensitive in preparations from dna C temperature-sensitive cells, and are both undetectable in preparations from dna D temperature-sensitive cells.

Keywords: ϕX174 DNA, DNA replication, DNA initiation

Full text

PDF

Selected References

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

  1. Campbell J. L., Soll L., Richardson C. C. Isolation and partial characterization of a mutant of Escherichia coli deficient in DNA polymerase II. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2090–2094. doi: 10.1073/pnas.69.8.2090. [DOI] [PMC free article] [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. Fuchs J. A., Karlström H. O., Warner H. R., Reichard P. Defective gene product in dnaF mutant of Escherichia coli. Nat New Biol. 1972 Jul 19;238(81):69–71. doi: 10.1038/newbio238069a0. [DOI] [PubMed] [Google Scholar]
  4. Gefter M. L., Hirota Y., Kornberg T., Wechsler J. A., Barnoux C. Analysis of DNA polymerases II and 3 in mutants of Escherichia coli thermosensitive for DNA synthesis. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3150–3153. doi: 10.1073/pnas.68.12.3150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hirota Y., Mordoh J., Jacob F. On the process of cellular division in Escherichia coli. 3. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. J Mol Biol. 1970 Nov 14;53(3):369–387. doi: 10.1016/0022-2836(70)90072-0. [DOI] [PubMed] [Google Scholar]
  6. Hirota Y., Ryter A., Jacob F. Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968;33:677–693. doi: 10.1101/sqb.1968.033.01.077. [DOI] [PubMed] [Google Scholar]
  7. Hurwitz J., Wickner S., Wright M. Studies on in vitro DNA synthesis. II. Isolation of a protein which stimulates deoxynucleotide incorporation catalyzed by DNA polymerase of E. coli. Biochem Biophys Res Commun. 1973 Mar 17;51(2):257–267. doi: 10.1016/0006-291x(73)91251-5. [DOI] [PubMed] [Google Scholar]
  8. Marinus M. G., Adelberg E. A. Vegetative Replication and Transfer Replication of Deoxyribonucleic Acid in Temperature-Sensitive Mutants of Escherichia coli K-12. J Bacteriol. 1970 Dec;104(3):1266–1272. doi: 10.1128/jb.104.3.1266-1272.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nüsslein V., Otto B., Bonhoeffer F., Schaller H. Function of DNA polymerase 3 in DNA replication. Nat New Biol. 1971 Dec 29;234(52):285–286. doi: 10.1038/newbio234285a0. [DOI] [PubMed] [Google Scholar]
  10. Schaller H., Nüsslein C., Bonhoeffer F. J., Kurz C., Nietzschmann I. Affinity chromatography of DNA-binding enzymes on single-stranded DNA-agarose columns. Eur J Biochem. 1972 Apr 24;26(4):474–481. doi: 10.1111/j.1432-1033.1972.tb01789.x. [DOI] [PubMed] [Google Scholar]
  11. Schekman R., Wickner W., Westergaard O., Brutlag D., Geider K., Bertsch L. L., Kornberg A. Initiation of DNA synthesis: synthesis of phiX174 replicative form requires RNA synthesis resistant to rifampicin. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2691–2695. doi: 10.1073/pnas.69.9.2691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Schubach W. H., Whitmer J. D., Davern C. I. Genetic control of DNA initiation in Escherichia coli. J Mol Biol. 1973 Feb 25;74(2):205–221. doi: 10.1016/0022-2836(73)90107-1. [DOI] [PubMed] [Google Scholar]
  13. Sigal N., Delius H., Kornberg T., Gefter M. L., Alberts B. A DNA-unwinding protein isolated from Escherichia coli: its interaction with DNA and with DNA polymerases. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3537–3541. doi: 10.1073/pnas.69.12.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Wickner R. B., Ginsberg B., Berkower I., Hurwitz J. Deoxyribonucleic acid plymerase II. of Escherichia coli. I. The purification and characterization of the enzyme. J Biol Chem. 1972 Jan 25;247(2):489–497. [PubMed] [Google Scholar]
  17. 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]
  18. Wickner W., Brutlag D., Schekman R., Kornberg A. RNA synthesis initiates in vitro conversion of M13 DNA to its replicative form. Proc Natl Acad Sci U S A. 1972 Apr;69(4):965–969. doi: 10.1073/pnas.69.4.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wolf B. The characteristics and genetic map location of a temperature sensitive DNA mutant of E. coli K12. Genetics. 1972 Dec;72(4):569–593. doi: 10.1093/genetics/72.4.569. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES