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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
. 1986 Mar;83(5):1256–1260. doi: 10.1073/pnas.83.5.1256

Cloning of the Escherichia coli gene for primosomal protein i: the relationship to dnaT, essential for chromosomal DNA replication.

H Masai, M W Bond, K Arai
PMCID: PMC323054  PMID: 3006041

Abstract

The Escherichia coli gene encoding one of the primosomal proteins, protein i, was cloned by the use of synthetic oligonucleotide probes. Nucleotide sequence analysis revealed a coding region for protein i of 537 base pairs preceded by a possible promoter sequence. The gene is located adjacent to the dnaC locus, probably both being in a single operon. The protein i gene was shown to be closely related to the dnaT locus based on the following observations. (i) A multicopy plasmid carrying only the protein i gene suppresses the temperature-sensitive phenotype of a dnaT strain and restores the ability of the strain to carry out stable DNA replication in the absence of protein synthesis. (ii) An extract from a dnaT strain does not support replication of the plasmid pBR322 in vitro; addition of purified protein i restores its activity. These results indicate that protein i is encoded by dnaT and that it is essential for chromosomal DNA replication and is involved in the induction of stable DNA replication during the SOS response.

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Selected References

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

  1. Arai K., Arai N., Shlomai J., Kornberg A. Replication of duplex DNA of phage phi X174 reconstituted with purified enzymes. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3322–3326. doi: 10.1073/pnas.77.6.3322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arai K., Kornberg A. Unique primed start of phage phi X174 DNA replication and mobility of the primosome in a direction opposite chain synthesis. Proc Natl Acad Sci U S A. 1981 Jan;78(1):69–73. doi: 10.1073/pnas.78.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arai K., Low R. L., Kornberg A. Movement and site selection for priming by the primosome in phage phi X174 DNA replication. Proc Natl Acad Sci U S A. 1981 Feb;78(2):707–711. doi: 10.1073/pnas.78.2.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arai K., Low R., Kobori J., Shlomai J., Kornberg A. Mechanism of dnaB protein action. V. Association of dnaB protein, protein n', and other repriming proteins in the primosome of DNA replication. J Biol Chem. 1981 May 25;256(10):5273–5280. [PubMed] [Google Scholar]
  5. Arai K., McMacken R., Yasuda S., Kornberg A. Purification and properties of Escherichia coli protein i, a prepriming protein in phi X174 DNA replication. J Biol Chem. 1981 May 25;256(10):5281–5286. [PubMed] [Google Scholar]
  6. Fuller R. S., Kaguni J. M., Kornberg A. Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7370–7374. doi: 10.1073/pnas.78.12.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Heidecker G., Messing J., Gronenborn B. A versatile primer for DNA sequencing in the M13mp2 cloning system. Gene. 1980 Jun;10(1):69–73. doi: 10.1016/0378-1119(80)90145-6. [DOI] [PubMed] [Google Scholar]
  8. Imber R., Low R. L., Ray D. S. Identification of a primosome assembly site in the region of the ori 2 replication origin of the Escherichia coli mini-F plasmid. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7132–7136. doi: 10.1073/pnas.80.23.7132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kobori J. A., Kornberg A. The Escherichia coli dnaC gene product. I. Overlapping of the dnaC proteins of Escherichia coli and Salmonella typhimurium by cloning into a high copy number plasmid. J Biol Chem. 1982 Nov 25;257(22):13757–13762. [PubMed] [Google Scholar]
  10. Kogoma T., Lark K. G. Characterization of the replication of Escherichia coli DNA in the absence of protein synthesis: stable DNA replication. J Mol Biol. 1975 May 15;94(2):243–256. doi: 10.1016/0022-2836(75)90081-9. [DOI] [PubMed] [Google Scholar]
  11. Lark C. A., Riazi J., Lark K. G. dnaT, dominant conditional-lethal mutation affecting DNA replication in Escherichia coli. J Bacteriol. 1978 Dec;136(3):1008–1017. doi: 10.1128/jb.136.3.1008-1017.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Low R. L., Arai K., Kornberg A. Conservation of the primosome in successive stages of phi X174 DNA replication. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1436–1440. doi: 10.1073/pnas.78.3.1436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Minden J. S., Marians K. J. Replication of pBR322 DNA in vitro with purified proteins. Requirement for topoisomerase I in the maintenance of template specificity. J Biol Chem. 1985 Aug 5;260(16):9316–9325. [PubMed] [Google Scholar]
  14. Nakayama N., Arai N., Bond M. W., Kaziro Y., Arai K. Nucleotide sequence of dnaB and the primary structure of the dnaB protein from Escherichia coli. J Biol Chem. 1984 Jan 10;259(1):97–101. [PubMed] [Google Scholar]
  15. Nomura N., Low R. L., Ray D. S. Identification of ColE1 DNA sequences that direct single strand-to-double strand conversion by a phi X174 type mechanism. Proc Natl Acad Sci U S A. 1982 May;79(10):3153–3157. doi: 10.1073/pnas.79.10.3153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pribnow D. Bacteriophage T7 early promoters: nucleotide sequences of two RNA polymerase binding sites. J Mol Biol. 1975 Dec 15;99(3):419–443. doi: 10.1016/s0022-2836(75)80136-7. [DOI] [PubMed] [Google Scholar]
  17. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shlomai J., Kornberg A. An Escherichia coli replication protein that recognizes a unique sequence within a hairpin region in phi X174 DNA. Proc Natl Acad Sci U S A. 1980 Feb;77(2):799–803. doi: 10.1073/pnas.77.2.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Zipursky S. L., Marians K. J. Identification of two Escherichia coli factor Y effector sites near the origins of replication of the plasmids (ColE1 and pBR322. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6521–6525. doi: 10.1073/pnas.77.11.6521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. van der Ende A., Teertstra R., van der Avoort H. G., Weisbeek P. J. Initiation signals for complementary strand DNA synthesis on single-stranded plasmid DNA. Nucleic Acids Res. 1983 Jul 25;11(14):4957–4975. doi: 10.1093/nar/11.14.4957. [DOI] [PMC free article] [PubMed] [Google Scholar]

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