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. 1989 Oct;171(10):5672–5679. doi: 10.1128/jb.171.10.5672-5679.1989

DNA replication in Escherichia coli mutants that lack protein HU.

T Ogawa 1, M Wada 1, Y Kano 1, F Imamoto 1, T Okazaki 1
PMCID: PMC210413  PMID: 2676987

Abstract

DNA replication in Escherichia coli cells lacking protein HU was studied. HU has been suggested to be involved in the initiation of replication from in vitro studies. The isolated HU mutants, however, are viable under normal growth conditions (M. Wada, Y. Kano, T. Ogawa, T. Okazaki, and F. Imamoto, J. Mol. Biol. 204:581-591, 1988). Chromosomal replication in the mutants appeared to be normal with respect to bidirectional replication from oriC and to its dependence on dnaA and some other dna gene products. No significant defect was observed in DNA synthesis in vitro with a crude enzyme fraction prepared from the mutant cells. These results, along with the earlier in vitro studies, suggest that other histonelike protein(s) may substitute for HU in the initiation of replication in the mutant cells. Minichromosomes were more unstable in the mutants. In the absence of either the mioC promoter, from which transcription enters oriC, or the DnaA box (DnaA protein-binding site) just upstream of the mioC promoter, the minichromosomes were especially unstable in the HU mutant and were integrated into the chromosomal oriC region under conditions selective for the plasmid-harboring cells.

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Selected References

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  1. Arai K., Yasuda S., Kornberg A. Mechanism of dnaB protein action. I. Crystallization and properties of dnaB protein, an essential replication protein in Escherichia coli. J Biol Chem. 1981 May 25;256(10):5247–5252. [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baker T. A., Kornberg A. Transcriptional activation of initiation of replication from the E. coli chromosomal origin: an RNA-DNA hybrid near oriC. Cell. 1988 Oct 7;55(1):113–123. doi: 10.1016/0092-8674(88)90014-1. [DOI] [PubMed] [Google Scholar]
  5. Baker T. A., Sekimizu K., Funnell B. E., Kornberg A. Extensive unwinding of the plasmid template during staged enzymatic initiation of DNA replication from the origin of the Escherichia coli chromosome. Cell. 1986 Apr 11;45(1):53–64. doi: 10.1016/0092-8674(86)90537-4. [DOI] [PubMed] [Google Scholar]
  6. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  7. Bramhill D., Kornberg A. Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome. Cell. 1988 Mar 11;52(5):743–755. doi: 10.1016/0092-8674(88)90412-6. [DOI] [PubMed] [Google Scholar]
  8. Broyles S. S., Pettijohn D. E. Interaction of the Escherichia coli HU protein with DNA. Evidence for formation of nucleosome-like structures with altered DNA helical pitch. J Mol Biol. 1986 Jan 5;187(1):47–60. doi: 10.1016/0022-2836(86)90405-5. [DOI] [PubMed] [Google Scholar]
  9. Buhk H. J., Messer W. The replication origin region of Escherichia coli: nucleotide sequence and functional units. Gene. 1983 Oct;24(2-3):265–279. doi: 10.1016/0378-1119(83)90087-2. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Craigie R., Arndt-Jovin D. J., Mizuuchi K. A defined system for the DNA strand-transfer reaction at the initiation of bacteriophage Mu transposition: protein and DNA substrate requirements. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7570–7574. doi: 10.1073/pnas.82.22.7570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dixon N. E., Kornberg A. Protein HU in the enzymatic replication of the chromosomal origin of Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jan;81(2):424–428. doi: 10.1073/pnas.81.2.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Drlica K., Rouviere-Yaniv J. Histonelike proteins of bacteria. Microbiol Rev. 1987 Sep;51(3):301–319. doi: 10.1128/mr.51.3.301-319.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  15. Flashner Y., Gralla J. D. DNA dynamic flexibility and protein recognition: differential stimulation by bacterial histone-like protein HU. Cell. 1988 Aug 26;54(5):713–721. doi: 10.1016/s0092-8674(88)80016-3. [DOI] [PubMed] [Google Scholar]
  16. Fuller R. S., Funnell B. E., Kornberg A. The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites. Cell. 1984 Oct;38(3):889–900. doi: 10.1016/0092-8674(84)90284-8. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Funnell B. E., Baker T. A., Kornberg A. In vitro assembly of a prepriming complex at the origin of the Escherichia coli chromosome. J Biol Chem. 1987 Jul 25;262(21):10327–10334. [PubMed] [Google Scholar]
  19. Hirota Y., Jacob F., Ryter A., Buttin G., Nakai T. On the process of cellular division in Escherichia coli. I. Asymmetrical cell division and production of deoxyribonucleic acid-less bacteria. J Mol Biol. 1968 Jul 14;35(1):175–192. doi: 10.1016/s0022-2836(68)80046-4. [DOI] [PubMed] [Google Scholar]
  20. Jaffé A., D'Ari R., Norris V. SOS-independent coupling between DNA replication and cell division in Escherichia coli. J Bacteriol. 1986 Jan;165(1):66–71. doi: 10.1128/jb.165.1.66-71.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Johnson R. C., Bruist M. F., Simon M. I. Host protein requirements for in vitro site-specific DNA inversion. Cell. 1986 Aug 15;46(4):531–539. doi: 10.1016/0092-8674(86)90878-0. [DOI] [PubMed] [Google Scholar]
  22. Junker D. E., Jr, Rokeach L. A., Ganea D., Chiaramello A., Zyskind J. W. Transcription termination within the Escherichia coli origin of DNA replication, oriC. Mol Gen Genet. 1986 Apr;203(1):101–109. doi: 10.1007/BF00330390. [DOI] [PubMed] [Google Scholar]
  23. Keller W. Determination of the number of superhelical turns in simian virus 40 DNA by gel electrophoresis. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4876–4880. doi: 10.1073/pnas.72.12.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kogoma T. A novel Escherichia coli mutant capable of DNA replication in the absence of protein synthesis. J Mol Biol. 1978 May 5;121(1):55–69. doi: 10.1016/0022-2836(78)90262-0. [DOI] [PubMed] [Google Scholar]
  25. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  26. Leonard A. C., Whitford W. G., Helmstetter C. E. Involvement of DNA superhelicity in minichromosome maintenance in Escherichia coli. J Bacteriol. 1985 Feb;161(2):687–695. doi: 10.1128/jb.161.2.687-695.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Liu L. F., Wang J. C. Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7024–7027. doi: 10.1073/pnas.84.20.7024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lother H., Kölling R., Kücherer C., Schauzu M. dnaA protein-regulated transcription: effects on the in vitro replication of Escherichia coli minichromosomes. EMBO J. 1985 Feb;4(2):555–560. doi: 10.1002/j.1460-2075.1985.tb03664.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lother H., Messer W. Promoters in the E. coli replication origin. Nature. 1981 Nov 26;294(5839):376–378. doi: 10.1038/294376a0. [DOI] [PubMed] [Google Scholar]
  30. Løbner-Olesen A., Atlung T., Rasmussen K. V. Stability and replication control of Escherichia coli minichromosomes. J Bacteriol. 1987 Jun;169(6):2835–2842. doi: 10.1128/jb.169.6.2835-2842.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Masters M., Andresdottir V., Wolf-Watz H. Plasmids carrying oriC can integrate at or near the chromosome origin of Escherichia coli in the absence of a functional recA product. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1069–1072. doi: 10.1101/sqb.1979.043.01.118. [DOI] [PubMed] [Google Scholar]
  32. Norris V., Alliotte T., Jaffé A., D'Ari R. DNA replication termination in Escherichia coli parB (a dnaG allele), parA, and gyrB mutants affected in DNA distribution. J Bacteriol. 1986 Nov;168(2):494–504. doi: 10.1128/jb.168.2.494-504.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nozaki N., Okazaki T., Ogawa T. In vitro transcription of the origin region of replication of the Escherichia coli chromosome. J Biol Chem. 1988 Oct 5;263(28):14176–14183. [PubMed] [Google Scholar]
  34. Ogawa T., Baker T. A., van der Ende A., Kornberg A. Initiation of enzymatic replication at the origin of the Escherichia coli chromosome: contributions of RNA polymerase and primase. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3562–3566. doi: 10.1073/pnas.82.11.3562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Orr E., Fairweather N. F., Holland I. B., Pritchard R. H. Isolation and characterisation of a strain carrying a conditional lethal mutation in the cou gene of Escherichia coli K12. Mol Gen Genet. 1979;177(1):103–112. doi: 10.1007/BF00267259. [DOI] [PubMed] [Google Scholar]
  36. Rouvière-Yaniv J., Gros F. Characterization of a novel, low-molecular-weight DNA-binding protein from Escherichia coli. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3428–3432. doi: 10.1073/pnas.72.9.3428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rouvière-Yaniv J., Yaniv M., Germond J. E. E. coli DNA binding protein HU forms nucleosomelike structure with circular double-stranded DNA. Cell. 1979 Jun;17(2):265–274. doi: 10.1016/0092-8674(79)90152-1. [DOI] [PubMed] [Google Scholar]
  38. Schauzu M. A., Kücherer C., Kölling R., Messer W., Lother H. Transcripts within the replication origin, oriC, of Escherichia coli. Nucleic Acids Res. 1987 Mar 25;15(6):2479–2497. doi: 10.1093/nar/15.6.2479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sekimizu K., Bramhill D., Kornberg A. ATP activates dnaA protein in initiating replication of plasmids bearing the origin of the E. coli chromosome. Cell. 1987 Jul 17;50(2):259–265. doi: 10.1016/0092-8674(87)90221-2. [DOI] [PubMed] [Google Scholar]
  40. Sekimizu K., Bramhill D., Kornberg A. Sequential early stages in the in vitro initiation of replication at the origin of the Escherichia coli chromosome. J Biol Chem. 1988 May 25;263(15):7124–7130. [PubMed] [Google Scholar]
  41. Stuitje A. R., Meijer M. Maintenance and incompatibility of plasmids carrying the replication origin of the Escherichia coli chromosome: evidence for a control region of replication between oriC and asnA. Nucleic Acids Res. 1983 Aug 25;11(16):5775–5791. doi: 10.1093/nar/11.16.5775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stuitje A. R., de Wind N., van der Spek J. C., Pors T. H., Meijer M. Dissection of promoter sequences involved in transcriptional activation of the Escherichia coli replication origin. Nucleic Acids Res. 1986 Mar 11;14(5):2333–2344. doi: 10.1093/nar/14.5.2333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sueoka N., Yoshikawa H. The chromosome of Bacillus subtilis. I. Theory of marker frequency analysis. Genetics. 1965 Oct;52(4):747–757. doi: 10.1093/genetics/52.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tanaka M., Hiraga S. Negative control of oriC plasmid replication by transcription of the oriC region. Mol Gen Genet. 1985;200(1):21–26. doi: 10.1007/BF00383307. [DOI] [PubMed] [Google Scholar]
  45. Wada M., Kano Y., Ogawa T., Okazaki T., Imamoto F. Construction and characterization of the deletion mutant of hupA and hupB genes in Escherichia coli. J Mol Biol. 1988 Dec 5;204(3):581–591. doi: 10.1016/0022-2836(88)90357-9. [DOI] [PubMed] [Google Scholar]
  46. Wang Q. P., Kaguni J. M. Transcriptional repression of the dnaA gene of Escherichia coli by dnaA protein. Mol Gen Genet. 1987 Oct;209(3):518–525. doi: 10.1007/BF00331158. [DOI] [PubMed] [Google Scholar]
  47. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]
  48. Yamazaki K., Nagata A., Kano Y., Imamoto F. Isolation and characterization of nucleoid proteins from Escherichia coli. Mol Gen Genet. 1984;196(2):217–224. doi: 10.1007/BF00328053. [DOI] [PubMed] [Google Scholar]
  49. de Massy B., Fayet O., Kogoma T. Multiple origin usage for DNA replication in sdrA(rnh) mutants of Escherichia coli K-12. Initiation in the absence of oriC. J Mol Biol. 1984 Sep 15;178(2):227–236. doi: 10.1016/0022-2836(84)90141-4. [DOI] [PubMed] [Google Scholar]
  50. de Massy B., Patte J., Louarn J. M., Bouché J. P. oriX: a new replication origin in E. coli. Cell. 1984 Jan;36(1):221–227. doi: 10.1016/0092-8674(84)90092-8. [DOI] [PubMed] [Google Scholar]
  51. de Wind N., Parren P., Stuitje A. R., Meijer M. Evidence for the involvement of the 16kD gene promoter in initiation of chromosomal replication of Escherichia coli strains carrying a B/r-derived replication origin. Nucleic Acids Res. 1987 Jun 25;15(12):4901–4914. doi: 10.1093/nar/15.12.4901. [DOI] [PMC free article] [PubMed] [Google Scholar]

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