Skip to main content
NCBI home page
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 Nov;70(11):3120–3124. doi: 10.1073/pnas.70.11.3120

Studies on In Vitro DNA Synthesis Isolation of dna B Gene Product from Escherichia coli*

Michel Wright 1, Sue Wickner 1, Jerard Hurwitz 1
PMCID: PMC427183  PMID: 4274130

Abstract

Formation of duplex DNA from ϕX174 single-stranded DNA by extracts of E. coli was previously shown to require the gene product of dna B. Using as an assay the stimulation of ϕX174 DNA-dependent synthesis in inactivated extracts of dna B temperature-sensitive cells, we purified the dna B gene product from wild-type E. coli as well as from a dna B temperature-sensitive mutant. The dna B temperature-sensitive gene product is more thermolabile than its wild-type counterpart.

Keywords: DNA replication, ϕX174 DNA

Full text

PDF
3120

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. 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]
  4. 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]
  5. 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]
  6. Siccardi A. G., Shapiro B. M. On the process of cellular division in Escherichia coli. IV. Altered protein composition and turnover of the membranes of thermosensitive mutants defective in chromosomal replication. J Mol Biol. 1971 Mar 28;56(3):475–490. doi: 10.1016/0022-2836(71)90395-0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Taketo A. Sensitivity of Escherichia coli to viral nucleic acid. VI. Capacity of dna mutants and DNA polymerase-less mutants for multiplication of phiA and phiX 174. Mol Gen Genet. 1973 Mar 27;122(1):15–22. doi: 10.1007/BF00337970. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. 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]
  13. Wickner S., Wright M., Hurwitz J. Studies on in vitro DNA synthesis. Purification of the dna G gene product from Escherichia coli. Proc Natl Acad Sci U S A. 1973 May;70(5):1613–1618. doi: 10.1073/pnas.70.5.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]

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