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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
. 1981 Sep;78(9):5391–5395. doi: 10.1073/pnas.78.9.5391

The dnaN gene codes for the beta subunit of DNA polymerase III holoenzyme of escherichia coli.

P M Burgers, A Kornberg, Y Sakakibara
PMCID: PMC348751  PMID: 6458041

Abstract

An Escherichia coli mutant, dnaN59, stops DNA synthesis promptly upon a shift to a high temperature; the wild-type dnaN gene carried in a transducing phage encodes a polypeptide of about 41,000 daltons [Sakakibara, Y. & Mizukami, T. (1980) Mol. Gen. Genet. 178, 541-553; Yuasa, S. & Sakakibara, Y. (1980) Mol. Gen. Genet. 180, 267-273]. We now find that the product of dnaN gene is the beta subunit of DNA polymerase III holoenzyme, the principal DNA synthetic multipolypeptide complex in E. coli. The conclusion is based on the following observations: (i) Extracts from dnaN59 cells were defective in phage phi X174 and G4 DNA synthesis after the mutant cells had been exposed to the increased temperature. (ii) The enzymatic defect was overcome by addition of purified beta subunit but not by other subunits of DNA polymerase III holoenzyme or by other replication proteins required for phi X174 DNA synthesis. (iii) Partially purified beta subunit from the dnaN mutant, unlike that from the wild type, was inactive in reconstituting the holoenzyme when mixed with the other purified subunits. (iv) Increased dosage of the dnaN gene provided by a plasmid carrying the gene raised cellular levels of the beta subunit 5- to 6-fold.

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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. 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]
  3. Hübscher U., Kornberg A. The delta subunit of Escherichia coli DNA polymerase III holoenzyme is the dnaX gene product. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6284–6288. doi: 10.1073/pnas.76.12.6284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Johanson K. O., McHenry C. S. Purification and characterization of the beta subunit of the DNA polymerase III holoenzyme of Escherichia coli. J Biol Chem. 1980 Nov 25;255(22):10984–10990. [PubMed] [Google Scholar]
  6. Kuwabara N., Uchida H. Functional cooperation of the dnaE and dnaN gene products in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5764–5767. doi: 10.1073/pnas.78.9.5764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. McHenry C. S., Crow W. DNA polymerase III of Escherichia coli. Purification and identification of subunits. J Biol Chem. 1979 Mar 10;254(5):1748–1753. [PubMed] [Google Scholar]
  8. 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]
  9. Sakakibara Y., Mizukami T. A temperature-sensitive Escherichia coli mutant defective in DNA replication: dnaN, a new gene adjacent to the dnaA gene. Mol Gen Genet. 1980;178(3):541–553. doi: 10.1007/BF00337859. [DOI] [PubMed] [Google Scholar]
  10. Sakakibara Y., Tsukano H., Sako T. Organization and transcription of the dnaA and dnaN genes of Escherichia coli. Gene. 1981 Jan-Feb;13(1):47–55. doi: 10.1016/0378-1119(81)90042-1. [DOI] [PubMed] [Google Scholar]
  11. Sako T., Sakakibara Y. Coordinate expression of Escherichia coli dnaA and dnaN genes. Mol Gen Genet. 1980;179(3):521–526. doi: 10.1007/BF00271741. [DOI] [PubMed] [Google Scholar]
  12. Ullrich A., Shine J., Chirgwin J., Pictet R., Tischer E., Rutter W. J., Goodman H. M. Rat insulin genes: construction of plasmids containing the coding sequences. Science. 1977 Jun 17;196(4296):1313–1319. doi: 10.1126/science.325648. [DOI] [PubMed] [Google Scholar]
  13. Yuasa S., Sakakibara Y. Identification of the dnaA and dnaN gene products of Escherichia coli. Mol Gen Genet. 1980;180(2):267–273. doi: 10.1007/BF00425838. [DOI] [PubMed] [Google Scholar]

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