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Proc Natl Acad Sci U S A. Sep 1975; 72(9): 3342–3346.
PMCID: PMC432988

Association of phiX174 DNA-dependent ATPase activity with an Escherichia coli protein, replication factor Y, required for in vitro synthesis of phiX174 DNA.

Abstract

phiX174 DNA-dependent DNA synthesis is catalyzed in vitro by the combination of at least 11 purified protein fractions: dnaB, dnaC(D), and dnaG gene products, DNA polymerase III, DNA elongation factors I and II, DNA binding protein, and replication factors W, X, Y, and Z. The reaction requires ATP, 4 dNTPs, and Mg+2 and is specific for phiX174 (or phiXahb) DNA. Purified replication factor Y contains phiX174 (or phiXahb) DNA-dependent ATPase (or dATPase) activity. The ATPase activity is poorly stimulated by other single-stranded DNA, by double-stranded DNA, or by RNA. The products of the phiX174 DNA-dependent ATPase activity of factor Y are Pi and ADP (or dADP). The association of phiX174 DNA-dependent ATPase activity with factor Y was shown in the following ways: (a) the two activities copurified with a constant ratio; (b) they comigrated on native polyacrylamide gel electrophoresis; (c) both activities were heat-inactivated at the same rate; and (d) both showed identical patterns of N-ethylmaleimide sensitivity.

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Selected References

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  • 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. [PubMed]
  • Hurwitz J, Wickner S. Involvement of two protein factors and ATP in in vitro DNA synthesis catalyzed by DNA polymerase 3 of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Jan;71(1):6–10. [PMC free article] [PubMed]
  • Geider K, Kornberg A. Conversion of the M13 viral single strand to the double-stranded replicative forms by purified proteins. J Biol Chem. 1974 Jul 10;249(13):3999–4005. [PubMed]
  • Bradley DE. A comparative study of some properties of the phi-X174 type bacteriophages. Can J Microbiol. 1970 Oct;16(10):965–971. [PubMed]
  • Bone DR, Dowel CE. A mutant of bacteriophage øX174 which infects E. coli K12 strains. I. Isolation and partial characterization of øXtB. Virology. 1973 Apr;52(2):319–329. [PubMed]
  • Godson GN. Evolution of phi-chi 174. Isolation of four new phi-chi-like phages and comparison with phi-chi 174. Virology. 1974 Mar;58(1):272–289. [PubMed]
  • Schekman R, Weiner A, Kornberg A. Multienzyme systems of DNA replication. Science. 1974 Dec 13;186(4168):987–993. [PubMed]
  • Vito CC, Primrose SB, Dowell CE. Growth of a capsid mutant of bacteriophage phi X174 in a temperature-sensitive strain of Escherichia coli. J Virol. 1975 Feb;15(2):281–287. [PMC free article] [PubMed]
  • Wickner S, Hurwitz J. Conversion of phiX174 viral DNA to double-stranded form by purified Escherichia coli proteins. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4120–4124. [PMC free article] [PubMed]
  • Gefter ML, Hirota Y, Kornberg T, Wechsler JA, 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • 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. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Sigal N, Delius H, Kornberg T, Gefter ML, 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. [PMC free article] [PubMed]
  • Molineux IJ, Friedman S, Gefter ML. Purification and properties of the Escherichia coli deoxyribonucleic acid-unwinding protein. Effects on deoxyribonucleic acid synthesis in vitro. J Biol Chem. 1974 Oct 10;249(19):6090–6098. [PubMed]
  • Weiner JH, Bertsch LL, Kornberg A. The deoxyribonucleic acid unwinding protein of Escherichia coli. Properties and functions in replication. J Biol Chem. 1975 Mar 25;250(6):1972–1980. [PubMed]
  • Wright M, Wickner S, Hurwitz J. Studies on in vitro DNA synthesis. Isolation of DNA B gene product from Escherichia coli. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3120–3124. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Livingston DM, Hinkle DC, Richardson CC. Deoxyribonucleic acid polymerase III of Escherichia coli. Purification and properties. J Biol Chem. 1975 Jan 25;250(2):461–469. [PubMed]
  • CONWAY TW, 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. [PMC free article] [PubMed]
  • JOVIN T, CHRAMBACH A, NAUGHTON MA. AN APPARATUS FOR PREPARATIVE TEMPERATURE-REGULATED POLYACRYLAMIDE GEL ELECTROPHORESIS. Anal Biochem. 1964 Nov;9:351–369. [PubMed]
  • Ebisuzaki K, Behme MT, Senior C, Shannon D, Dunn D. An alternative approach to the study of new enzymatic reactions involving DNA (DNA-dependent ATPases-purification-properties-E. coli). Proc Natl Acad Sci U S A. 1972 Feb;69(2):515–519. [PMC free article] [PubMed]
  • Buttin G, Wright M. Enzymatic DNA degradation in E. coli: its relationship to synthetic processes at the chromosome level. Cold Spring Harb Symp Quant Biol. 1968;33:259–269. [PubMed]

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