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. 1995 Nov;177(22):6352–6361. doi: 10.1128/jb.177.22.6352-6361.1995

Lethality and survival of Klebsiella oxytoca evoked by conjugative IncN group plasmids.

M Rodríguez 1, M Holcík 1, V N Iyer 1
PMCID: PMC177484  PMID: 7592409

Abstract

The transmission of plasmid pCU1 (or other IncN group plasmid) into a population of Klebsiella oxytoca cells reduces the viability of the population. A 2,400-bp region adjacent to traA is responsible for this phenotype and includes two regions, called kikA and kikC. Klebsiella cells which received this region and survived were found to acquire a chromosomal mutation which renders them immune to killing even after the plasmid is cured from the cells. To obtain insight into the mode of this apparent lethality, an appropriate pCU1lacZ derivative was constructed. It could be introduced with high efficiency into Klebsiella cells. Analyses of the resultant colonies indicate that the loss of viability is not a consequence of the death of plasmid-free segregants. On the contrary and unlike postsegregational killing by plasmids, cells survived by losing the plasmid or by acquiring, secondarily, a chromosomal mutation which confers immunity to killing.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Figurski D. H., Pohlman R. F., Bechhofer D. H., Prince A. S., Kelton C. A. Broad host range plasmid RK2 encodes multiple kil genes potentially lethal to Escherichia coli host cells. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1935–1939. doi: 10.1073/pnas.79.6.1935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fürste J. P., Pansegrau W., Frank R., Blöcker H., Scholz P., Bagdasarian M., Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48(1):119–131. doi: 10.1016/0378-1119(86)90358-6. [DOI] [PubMed] [Google Scholar]
  8. Gerdes K., Poulsen L. K., Thisted T., Nielsen A. K., Martinussen J., Andreasen P. H. The hok killer gene family in gram-negative bacteria. New Biol. 1990 Nov;2(11):946–956. [PubMed] [Google Scholar]
  9. Grant S. G., Jessee J., Bloom F. R., Hanahan D. Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4645–4649. doi: 10.1073/pnas.87.12.4645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hengen P. N., Denicourt D., Iyer V. N. Isolation and characterization of kikA, a region on IncN group plasmids that determines killing of Klebsiella oxytoca. J Bacteriol. 1992 May;174(9):3070–3077. doi: 10.1128/jb.174.9.3070-3077.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hengen P. N., Iyer V. N. DNA cassettes containing the origin of transfer (oriT) of two broad-host-range transfer systems. Biotechniques. 1992 Jul;13(1):56-8, 60, 62. [PubMed] [Google Scholar]
  12. Herrero M., de Lorenzo V., Timmis K. N. Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria. J Bacteriol. 1990 Nov;172(11):6557–6567. doi: 10.1128/jb.172.11.6557-6567.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  14. Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
  15. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. doi: 10.1016/0378-1119(88)90117-5. [DOI] [PubMed] [Google Scholar]
  16. Konarska-Kozlowska M., Iyer V. N. Physical and genetic organization of the IncN-group plasmid pCU1. Gene. 1981 Aug;14(3):195–204. doi: 10.1016/0378-1119(81)90115-3. [DOI] [PubMed] [Google Scholar]
  17. Konarska-Kozlowska M., Iyer V. N. Sequence homology between Inc N group plasmids. Plasmid. 1983 Nov;10(3):211–223. doi: 10.1016/0147-619x(83)90035-5. [DOI] [PubMed] [Google Scholar]
  18. Lehnherr H., Maguin E., Jafri S., Yarmolinsky M. B. Plasmid addiction genes of bacteriophage P1: doc, which causes cell death on curing of prophage, and phd, which prevents host death when prophage is retained. J Mol Biol. 1993 Oct 5;233(3):414–428. doi: 10.1006/jmbi.1993.1521. [DOI] [PubMed] [Google Scholar]
  19. MacNeil D., Zhu J., Brill W. J. Regulation of nitrogen fixation in Klebsiella pneumoniae: isolation and characterization of strains with nif-lac fusions. J Bacteriol. 1981 Jan;145(1):348–357. doi: 10.1128/jb.145.1.348-357.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McMorrow I., Chin D. T., Fiebig K., Pierce J. L., Wilson D. M., Reeve E. C., Wilson T. H. The lactose carrier of Klebsiella pneumoniae M5a1; the physiology of transport and the nucleotide sequence of the lacY gene. Biochim Biophys Acta. 1988 Nov 22;945(2):315–323. doi: 10.1016/0005-2736(88)90494-4. [DOI] [PubMed] [Google Scholar]
  21. Meade H. M., Long S. R., Ruvkun G. B., Brown S. E., Ausubel F. M. Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis. J Bacteriol. 1982 Jan;149(1):114–122. doi: 10.1128/jb.149.1.114-122.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ogura T., Hiraga S. Mini-F plasmid genes that couple host cell division to plasmid proliferation. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4784–4788. doi: 10.1073/pnas.80.15.4784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pohlman R. F., Genetti H. D., Winans S. C. Common ancestry between IncN conjugal transfer genes and macromolecular export systems of plant and animal pathogens. Mol Microbiol. 1994 Nov;14(4):655–668. doi: 10.1111/j.1365-2958.1994.tb01304.x. [DOI] [PubMed] [Google Scholar]
  25. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  26. Rodriguez M., Iyer V. N. Killing of Klebsiella pneumoniae mediated by conjugation with bacteria carrying antibiotic-resistance plasmids of the group N. Plasmid. 1981 Jul;6(1):141–147. doi: 10.1016/0147-619x(81)90060-3. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schaffer H. E., Sederoff R. R. Improved estimation of DNA fragment lengths from Agarose gels. Anal Biochem. 1981 Jul 15;115(1):113–122. doi: 10.1016/0003-2697(81)90533-9. [DOI] [PubMed] [Google Scholar]
  29. Thatte V., Bradley D. E., Iyer V. N. N conjugative transfer system of plasmid pCU1. J Bacteriol. 1985 Sep;163(3):1229–1236. doi: 10.1128/jb.163.3.1229-1236.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Thatte V., Gill S., Iyer V. N. Regions on plasmid pCU1 required for the killing of Klebsiella pneumoniae. J Bacteriol. 1985 Sep;163(3):1296–1299. doi: 10.1128/jb.163.3.1296-1299.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tsuchimoto S., Nishimura Y., Ohtsubo E. The stable maintenance system pem of plasmid R100: degradation of PemI protein may allow PemK protein to inhibit cell growth. J Bacteriol. 1992 Jul;174(13):4205–4211. doi: 10.1128/jb.174.13.4205-4211.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wilmes-Riesenberg M. R., Wanner B. L. TnphoA and TnphoA' elements for making and switching fusions for study of transcription, translation, and cell surface localization. J Bacteriol. 1992 Jul;174(14):4558–4575. doi: 10.1128/jb.174.14.4558-4575.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Winans S. C., Walker G. C. Identification of pKM101-encoded loci specifying potentially lethal gene products. J Bacteriol. 1985 Jan;161(1):417–424. doi: 10.1128/jb.161.1.417-424.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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