Molecular and General Genetics MGG

, Volume 201, Issue 3, pp 467–476 | Cite as

Characterization and properties of very large inversions of the E. coli chromosome along the origin-to-terminus axis

  • J. M. Louarn
  • J. P. Bouché
  • F. Legendre
  • J. Louarn
  • J. Patte
Article

Summary

Suppression of a dnaA46 mutation by integration of plasmid R100.1 derivatives in the termination region of chromosome replication in E. coli results in medium dependence, the suppressed bacteria being sensitive to rich medium at 42° C. Derivatives of such bacteria have been selected for growth at 42° C in rich medium and we have analyzed representatives of the most frequently observed type: bacteria displaying, once cured of the suppressor plasmid, both rich-medium sensitivity and temperature sensitivity. We found, in all cases, that the chromosome had undergone a major inversion event between two inverted IS5's. One is located at 29.2 min on the chromosome map and the other at either one of two positions between 69 and 80 min. The consequences of such inversions for cell growth are discussed. Some of them result from the fact that the replication terminator T2 is located, in inverted chromosomes, close to oriC in the orientation which allows its functioning as a terminus (de Massy et al. in press). Our observations allow an estimation of the frequency of inversions arising from recombination between pairs of inverted chromosomal IS, which could be as high as 10-2 per cell per generation. We also found that inversion reversal occurs frequently after Hfr conjugational transfer of one of the IS5's, in its wild-type location. This led us to propose a new mechanisms of recombination, in which the incoming DNA strands serve as guides to favor recombination between the resident sequences.

Abbreviations

Sin

suppresive integration

Rms/Rmr

rich medium sensitivity/resistance

Ts/Tr

temperature sensitivity/resistance

Apr

ampicillin resistance

Nalr

nalidixic acid resistance

Spr

spectinomycin resistance

Str

streptomycin resistance

Tcr

tetracycline resistance

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References

  1. Armstrong K, Helinski RD (1977) In: Bukhari AI, Shapiro JA, Adhya SL (eds) DNA insertion elements, plasmids and episomes. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  2. Bachman BJ (1983) Linkage map of Escherichia coli K12. Edition 7. Microbiol Rev 47:180–230Google Scholar
  3. Béjar S, Bouché JP (1983) Molecular cloning of the region of the terminus of Escherichia coli K12 DNA replication. J Bacteriol 153:604–609Google Scholar
  4. Bouché JP (1982) Physical map of a 470 kilobase pair region flanking the terminus of DNA replication in the Escherichia coli K12 genome. J Mol Biol 154:1–20Google Scholar
  5. Bouché JP, Gélugne JP, Louarn J, Louarn JM, Kaiser K (1982) Relationships between the physical and genetic maps of a 470×103 base-pair region around the terminus of Escherichia coli K12 DNA replication. J Mol Biol 154:21–32Google Scholar
  6. Bouck N, Adelberg EA (1970) Mechanism of action of nalidixic acid on conjugating bacteria. J Bacteriol 102:688–701Google Scholar
  7. Caspers B, Dalrymple B, Iida S, Arber W (1984) IS30, a new insertion sequence of Escherichia coli. Mol gen Genet 196:68–73Google Scholar
  8. Casse F, Pascal M, Chippaux M (1973) Comparison between the genetic maps of Esherichia coli and Salmonella typhimurium: length of the inverted segment in the trp region. Mol Gen Genet 124:253–257Google Scholar
  9. de Massy B, Patte J, Louarn JM, Bouché JP (1984) oriX: a new replication origin in E. coli. Cell 36:221–227Google Scholar
  10. de Massy B, Béjar S, Louarn JM, Bouché JP (1985) The replication of integratively suppressed dnaA46 mutants of Escherichia coli. Possible significance of changes in fork velocity for nucleoid structure and evidence for two termini. J Mol Biol, in pressGoogle Scholar
  11. Haziza C, Stragier P, Patte JC (1982) Nucleotide sequence of the asd gene of Escherichia coli: absence of a typical attenuation signal. EMBO J 1:379–384Google Scholar
  12. Hill CW, Harnish BW (1981) Inversions between ribosomal RNA genes of Escherichia coli. Proc Natl Acad Sci USA 78:7069–7072Google Scholar
  13. Iida S, Meyer I, Arber W (1983) Prokaryotic IS elements. In: Shapiro JA (ed) Mobile genetic elements. Academic Press Inc, New York, pp 159–221Google Scholar
  14. Louarn J, Patte J, Louarn JM (1977) Evidence for a fixed termination site of chromosome replication in Escherichia coli K12. J Mol Biol 115:295–314Google Scholar
  15. Louarn J, Patte J, Louarn JM (1982) Suppression of Escherichia coli dna A46 mutations by integration of plasmid R100.1 derivatives: constraints imposed by the terminus. J Bacteriol 151:657–667Google Scholar
  16. Louarn J, Legrand P, Patte J, Louarn JM (1983) The terminus of chromosome replication of E. coli. Phenotypic suppression of a dnaA mutation by plasmid integration near terC. In: de Recondo AM (ed) New approaches in eukaryotic DNA replication. Plenum Press, New York, pp 247–263Google Scholar
  17. Low KB (1972) Escherichia coli K12 F-prime factors, old and new. Bacteriol Rev 36:587–607Google Scholar
  18. Low KB (1973) Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K12. J Bacteriol 113:798–812Google Scholar
  19. Maloy SR, Nunn WD (1981) Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol 145:1110–1112Google Scholar
  20. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  21. Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  22. Okita TW, Rodriguez RL, Preiss J (1981) Biosynthesis of bacterial glycogen. Cloning of the glycogen biosynthetic enzyme structural genes of Escherichia coli. JBC 256:6944–6949Google Scholar
  23. Prentki P, Krisch HM (1984) In-vitro insertional mutagenesis with a selectable DNA fragment. Gene 29:303–313Google Scholar
  24. Raibaud O, Schwartz M (1980) Restriction map of the Escherichia coli malA region and identification of the malT product. J Bacteriol 143:761–771Google Scholar
  25. Schmid MB, Roth JR (1983) Selection and endpoints distribution of bacterial inversion mutations. Genetics 105:539–557Google Scholar
  26. Schweizer H, Boos W (1984) Characterization of the ugp region containing the genes for the phoB dependent sn-glycerol-3-phosphate transport system of Escherichia coli. Mol Gen Genet 197:161–169Google Scholar
  27. Tresguerres EF, Nandadasa HG, Pritchard RH (1975) Suppression of initiation-negative strains of Escherichia coli by integration of the sex factor F. J Bacteriol 121:554–561Google Scholar
  28. Virolle MJ, Gélugne JP, Béjar S, Bouché JP (1983) Origin of Escherichia coli K12 Hfr B7. J Bacteriol 153:610–615Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • J. M. Louarn
    • 1
    • 2
  • J. P. Bouché
    • 1
    • 2
  • F. Legendre
    • 1
    • 2
  • J. Louarn
    • 1
    • 2
  • J. Patte
    • 1
    • 2
  1. 1.Centre de Biochimie et de Génétique cellulaires du C.N.R.S.Toulouse CedexFrance
  2. 2.Université Paul SabatierToulouse CedexFrance

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