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. 1987 Oct 12;15(19):7663–7675. doi: 10.1093/nar/15.19.7663

Relation of the Escherichia coli dnaX gene to its two products--the tau and gamma subunits of DNA polymerase III holoenzyme.

S H Lee 1, P Kanda 1, R C Kennedy 1, J R Walker 1
PMCID: PMC306299  PMID: 3313272

Abstract

The Escherichia coli DNA polymerase III holoenzyme 71.1 kDa tau subunit is a 643 amino acid protein encoded by the dnaX gene. This gene also encodes the holoenzyme 56.5 kDa gamma subunit. The tau factor (as a tau'-LacZ' fusion protein) has been isolated and shown to be cleaved in vitro to form gamma and a 135 kda C-terminal cleavage product. The tau'-LacZ' fusion protein, gamma, and the C-terminal cleavage product have been isolated. N-terminal sequencing has demonstrated that tau and gamma share the same N-terminal sequences and that tau is proteolytically cleaved in vitro between residues 498 and 499 to form gamma. In addition, residues 420-440 were shown to be present in both tau and gamma by use of antibody specific for a synthetic peptide corresponding to that sequence. Some mechanism functions in vivo to ensure that tau and gamma are synthesized in a ratio of about one-to-one, as shown by radioimmune precipitation of tau and gamma from cellular extracts.

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Selected References

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

  1. Biswas S. B., Kornberg A. Nucleoside triphosphate binding to DNA polymerase III holoenzyme of Escherichia coli. A direct photoaffinity labeling study. J Biol Chem. 1984 Jun 25;259(12):7990–7993. [PubMed] [Google Scholar]
  2. Burgers P. M., Kornberg A. ATP activation of DNA polymerase III holoenzyme from Escherichia coli. II. Initiation complex: stoichiometry and reactivity. J Biol Chem. 1982 Oct 10;257(19):11474–11478. [PubMed] [Google Scholar]
  3. Chanh T. C., Dreesman G. R., Kanda P., Linette G. P., Sparrow J. T., Ho D. D., Kennedy R. C. Induction of anti-HIV neutralizing antibodies by synthetic peptides. EMBO J. 1986 Nov;5(11):3065–3071. doi: 10.1002/j.1460-2075.1986.tb04607.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DiFrancesco R., Bhatnagar S. K., Brown A., Bessman M. J. The interaction of DNA polymerase III and the product of the Escherichia coli mutator gene, mutD. J Biol Chem. 1984 May 10;259(9):5567–5573. [PubMed] [Google Scholar]
  5. Echols H., Lu C., Burgers P. M. Mutator strains of Escherichia coli, mutD and dnaQ, with defective exonucleolytic editing by DNA polymerase III holoenzyme. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2189–2192. doi: 10.1073/pnas.80.8.2189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fay P. J., Johanson K. O., McHenry C. S., Bambara R. A. Size classes of products synthesized processively by DNA polymerase III and DNA polymerase III holoenzyme of Escherichia coli. J Biol Chem. 1981 Jan 25;256(2):976–983. [PubMed] [Google Scholar]
  7. Fay P. J., Johanson K. O., McHenry C. S., Bambara R. A. Size classes of products synthesized processively by two subassemblies of Escherichia coli DNA polymerase III holoenzyme. J Biol Chem. 1982 May 25;257(10):5692–5699. [PubMed] [Google Scholar]
  8. Fersht A. R., Knill-Jones J. W. Contribution of 3' leads to 5' exonuclease activity of DNA polymerase III holoenzyme from Escherichia coli to specificity. J Mol Biol. 1983 Apr 25;165(4):669–682. doi: 10.1016/s0022-2836(83)80273-3. [DOI] [PubMed] [Google Scholar]
  9. Flower A. M., McHenry C. S. The adjacent dnaZ and dnaX genes of Escherichia coli are contained within one continuous open reading frame. Nucleic Acids Res. 1986 Oct 24;14(20):8091–8101. doi: 10.1093/nar/14.20.8091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fowler A. V., Zabin I. The amino acid sequence of beta-galactosidase of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1507–1510. doi: 10.1073/pnas.74.4.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fry D. C., Kuby S. A., Mildvan A. S. ATP-binding site of adenylate kinase: mechanistic implications of its homology with ras-encoded p21, F1-ATPase, and other nucleotide-binding proteins. Proc Natl Acad Sci U S A. 1986 Feb;83(4):907–911. doi: 10.1073/pnas.83.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HOWARD-FLANDERS P., SIMSON E., THERIOT L. A LOCUS THAT CONTROLS FILAMENT FORMATION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI K-12. Genetics. 1964 Feb;49:237–246. doi: 10.1093/genetics/49.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Henson J. M., Chu H., Irwin C. A., Walker J. R. Isolation and characterization of dnaX and dnaY temperature-sensitive mutants of Escherichia coli. Genetics. 1979 Aug;92(4):1041–1059. doi: 10.1093/genetics/92.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Higgins C. F., Hiles I. D., Salmond G. P., Gill D. R., Downie J. A., Evans I. J., Holland I. B., Gray L., Buckel S. D., Bell A. W. A family of related ATP-binding subunits coupled to many distinct biological processes in bacteria. Nature. 1986 Oct 2;323(6087):448–450. doi: 10.1038/323448a0. [DOI] [PubMed] [Google Scholar]
  15. 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. doi: 10.1073/pnas.71.1.6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hübscher U., Kornberg A. The dnaZ protein, the gamma subunit of DNA polymerase III holoenzyme of Escherichia coli. J Biol Chem. 1980 Dec 25;255(24):11698–11703. [PubMed] [Google Scholar]
  17. Kennedy R. C., Henkel R. D., Pauletti D., Allan J. S., Lee T. H., Essex M., Dreesman G. R. Antiserum to a synthetic peptide recognizes the HTLV-III envelope glycoprotein. Science. 1986 Mar 28;231(4745):1556–1559. doi: 10.1126/science.3006246. [DOI] [PubMed] [Google Scholar]
  18. Kodaira M., Biswas S. B., Kornberg A. The dnaX gene encodes the DNA polymerase III holoenzyme tau subunit, precursor of the gamma subunit, the dnaZ gene product. Mol Gen Genet. 1983;192(1-2):80–86. doi: 10.1007/BF00327650. [DOI] [PubMed] [Google Scholar]
  19. Kwoh T. J., Chan P. T., Patrick M. H. Examination of newly synthesized DNA in Escherichia coli. Mol Gen Genet. 1979 May 23;173(1):85–93. doi: 10.1007/BF00267693. [DOI] [PubMed] [Google Scholar]
  20. Lanford R. E., Kanda P., Kennedy R. C. Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal. Cell. 1986 Aug 15;46(4):575–582. doi: 10.1016/0092-8674(86)90883-4. [DOI] [PubMed] [Google Scholar]
  21. Lee S. H., Walker J. R. Escherichia coli DnaX product, the tau subunit of DNA polymerase III, is a multifunctional protein with single-stranded DNA-dependent ATPase activity. Proc Natl Acad Sci U S A. 1987 May;84(9):2713–2717. doi: 10.1073/pnas.84.9.2713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maki H., Kornberg A. The polymerase subunit of DNA polymerase III of Escherichia coli. II. Purification of the alpha subunit, devoid of nuclease activities. J Biol Chem. 1985 Oct 25;260(24):12987–12992. [PubMed] [Google Scholar]
  23. McHenry C. S. DNA polymerase III holoenzyme of Escherichia coli: components and function of a true replicative complex. Mol Cell Biochem. 1985 Feb;66(1):71–85. doi: 10.1007/BF00231826. [DOI] [PubMed] [Google Scholar]
  24. McHenry C. S. Purification and characterization of DNA polymerase III'. Identification of tau as a subunit of the DNA polymerase III holoenzyme. J Biol Chem. 1982 Mar 10;257(5):2657–2663. [PubMed] [Google Scholar]
  25. McHenry C., Kornberg A. DNA polymerase III holoenzyme of Escherichia coli. Purification and resolution into subunits. J Biol Chem. 1977 Sep 25;252(18):6478–6484. [PubMed] [Google Scholar]
  26. Mullin D. A., Woldringh C. L., Henson J. M., Walker J. R. Cloning of the Escherichia coli dnaZX region and identification of its products. Mol Gen Genet. 1983;192(1-2):73–79. doi: 10.1007/BF00327649. [DOI] [PubMed] [Google Scholar]
  27. O'Donnell M. E., Kornberg A. Complete replication of templates by Escherichia coli DNA polymerase III holoenzyme. J Biol Chem. 1985 Oct 15;260(23):12884–12889. [PubMed] [Google Scholar]
  28. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  29. Pauletti D., Simmonds R., Dreesman G. R., Kennedy R. C. Application of a modified computer algorithm in determining potential antigenic determinants associated with the AIDS virus glycoprotein. Anal Biochem. 1985 Dec;151(2):540–546. doi: 10.1016/0003-2697(85)90217-9. [DOI] [PubMed] [Google Scholar]
  30. Ryals J., Little R., Bremer H. Temperature dependence of RNA synthesis parameters in Escherichia coli. J Bacteriol. 1982 Aug;151(2):879–887. doi: 10.1128/jb.151.2.879-887.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Scheuermann R. H., Echols H. A separate editing exonuclease for DNA replication: the epsilon subunit of Escherichia coli DNA polymerase III holoenzyme. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7747–7751. doi: 10.1073/pnas.81.24.7747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Scheuermann R., Tam S., Burgers P. M., Lu C., Echols H. Identification of the epsilon-subunit of Escherichia coli DNA polymerase III holoenzyme as the dnaQ gene product: a fidelity subunit for DNA replication. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7085–7089. doi: 10.1073/pnas.80.23.7085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Spanos A., Sedgwick S. G., Yarranton G. T., Hübscher U., Banks G. R. Detection of the catalytic activities of DNA polymerases and their associated exonucleases following SDS-polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Apr 24;9(8):1825–1839. doi: 10.1093/nar/9.8.1825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Welch M. M., McHenry C. S. Cloning and identification of the product of the dnaE gene of Escherichia coli. J Bacteriol. 1982 Oct;152(1):351–356. doi: 10.1128/jb.152.1.351-356.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wickner S. H., Wickner R. B., Raetz C. R. Overproduction of DNA gene products by Escherichia coli strains carrying hybrid ColE1 plasmids. Biochem Biophys Res Commun. 1976 May 17;70(2):389–396. doi: 10.1016/0006-291x(76)91058-5. [DOI] [PubMed] [Google Scholar]
  37. Wickner S., Hurwitz J. Involvement of escherichia coli dnaZ gene product in DNA elongation in vitro. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1053–1057. doi: 10.1073/pnas.73.4.1053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wickner S. Mechanism of DNA elongation catalyzed by Escherichia coli DNA polymerase III, dnaZ protein, and DNA elongation factors I and III. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3511–3515. doi: 10.1073/pnas.73.10.3511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wickner W., Kornberg A. A holoenzyme form of deoxyribonucleic acid polymerase III. Isolation and properties. J Biol Chem. 1974 Oct 10;249(19):6244–6249. [PubMed] [Google Scholar]
  40. 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]
  41. Yasuda S., Takagi T. Overproduction of Escherichia coli replication proteins by the use of runaway-replication plasmids. J Bacteriol. 1983 Jun;154(3):1153–1161. doi: 10.1128/jb.154.3.1153-1161.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yin K. C., Blinkowa A., Walker J. R. Nucleotide sequence of the Escherichia coli replication gene dnaZX. Nucleic Acids Res. 1986 Aug 26;14(16):6541–6549. doi: 10.1093/nar/14.16.6541. [DOI] [PMC free article] [PubMed] [Google Scholar]

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