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. 1974 Jan;71(1):6–10. doi: 10.1073/pnas.71.1.6

Involvement of Two Protein Factors and ATP in In Vitro DNA Synthesis Catalyzed by DNA Polymerase III of Escherichia coli

Jerard Hurwitz 1, Sue Wickner 1
PMCID: PMC387920  PMID: 4589895

Abstract

The in vitro conversion of single-stranded DNA from bacteriophage fd to duplex structures depends on E. coli RNA polymerase, DNA polymerase III, riboand deoxyribonucleoside triphosphates, Mg+2, spermidine or DNA-unwinding protein of E. coli, and two additional protein factors, referred to here as Factors I and II. These two factors are also essential for dTMP incorporation catalyzed by DNA polymerase III and dependent on poly(dA)·oligo(dT) primer-template. In the latter reaction, there is an absolute dependency on ATP or dATP.

Keywords: DNA replication, fd DNA, poly(dA)·oligo(dT)

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Selected References

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

  1. Barry J., Alberts B. In vitro complementation as an assay for new proteins required for bacteriophage T4 DNA replication: purification of the complex specified by T4 genes 44 and 62. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2717–2721. doi: 10.1073/pnas.69.9.2717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CONWAY T. W., LIPMANN F. CHARACTERIZATION OF A RIBOSOME-LINKED GUANOSINE TRIPHOSPHATASE IN ESCHERICHIA COLI EXTRACTS. Proc Natl Acad Sci U S A. 1964 Dec;52:1462–1469. doi: 10.1073/pnas.52.6.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. 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]
  7. Hurwitz J., Yarbrough L., Wickner S. Utilization of deoxynucleoside triphosphates by DNA-dependent RNA polymerase of E. coli. Biochem Biophys Res Commun. 1972 Aug 7;48(3):628–635. doi: 10.1016/0006-291x(72)90394-4. [DOI] [PubMed] [Google Scholar]
  8. JOVIN T., CHRAMBACH A., NAUGHTON M. A. AN APPARATUS FOR PREPARATIVE TEMPERATURE-REGULATED POLYACRYLAMIDE GEL ELECTROPHORESIS. Anal Biochem. 1964 Nov;9:351–369. doi: 10.1016/0003-2697(64)90192-7. [DOI] [PubMed] [Google Scholar]
  9. Kornberg T., Gefter M. L. Deoxyribonucleic acid synthesis in cell-free extracts. IV. Purification and catalytic properties of deoxyribonucleic acid polymerase III. J Biol Chem. 1972 Sep 10;247(17):5369–5375. [PubMed] [Google Scholar]
  10. Moses R. E., Richardson C. C. Replication and repair of DNA in cells of Escherichia coli treated with toluene. Proc Natl Acad Sci U S A. 1970 Oct;67(2):674–681. doi: 10.1073/pnas.67.2.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Otto B., Bonhoeffer F., Schaller H. Purification and properties of DNA polymerase 3. Eur J Biochem. 1973 May 2;34(3):440–447. doi: 10.1111/j.1432-1033.1973.tb02777.x. [DOI] [PubMed] [Google Scholar]
  13. Pisetsky D., Berkower I., Wickner R., Hurwitz J. Role of ATP in DNA synthesis in Escherichia coli. J Mol Biol. 1972 Nov 28;71(3):557–571. doi: 10.1016/s0022-2836(72)80023-8. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Vosberg H. P., Hoffmann-Berling H. DNA synthesis in nucleotide-permeable Escherichia coli cells. I. Preparation and properties of ether-treated cells. J Mol Biol. 1971 Jun 28;58(3):739–753. doi: 10.1016/0022-2836(71)90037-4. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Wickner R. B., Hurwitz J. DNA replication in Escherichia coli made permeable by treatment with high sucrose. Biochem Biophys Res Commun. 1972 Apr 14;47(1):202–211. doi: 10.1016/s0006-291x(72)80029-9. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]
  23. Wickner W., Schekman R., Geider K., Kornberg A. A new form of DNA polymerase 3 and a copolymerase replicate a long, single-stranded primer-template. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1764–1767. doi: 10.1073/pnas.70.6.1764. [DOI] [PMC free article] [PubMed] [Google Scholar]

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