Summary
Clo DF13 plasmids that are present at high copy-number in bacterial cells, such as Clo DF13 cop1 Ts, cop2 and cop3 are not stably inherited in the progeny, when certain plasmid DNA regions have been deleted. We have localized two Clo DF13 DNA regions involved in stable maintenance through accurate partitioning (par) namely parA, located between 71% and 72% and parB, located between 45% and 50% on the Clo DF13 genome. The instability of these cop plasmids which is accompanied by the formation of high amounts of multimeric DNA molecules, could be abolished by the insertion of transposon Tn901 into the plasmid genome. In particular that part of Tn901, that encodes for the site-specific recombination/ resolution system, appeared to be essential for stabilizing plasmid molecules. Wild-type parA- and/or parB- Clo DF13 plasmids, in contrast to cop mutants lacking these regions, are stably maintained during subsequent cell division, indicating that other (host specified) functions contribute to plasmid stability. Analysis of the role of host recombination systems in plasmid partitioning revealed that the recA function has no influence and recBC contributes only weakly to plasmid stability. With respect to the recE pathway, however, we found that in a recE proficient host all plasmids, even those lacking parA and/or parB, are stably maintained, indicating that the function of parA and parB can be replaced not only by the site-specific resolution functions of transposon Tn901, but also by the recE system. The possible role of plasmid specified and host specified functions in plasmid partitioning will be discussed.
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References
Adler HI, Fisher WD, Cohen A, Hardigree AA (1967) Miniature Escherichia coli cells deficient in DNA Proc Natl Acad Sci USA 57:321–326
Andreoli PM, Brandsma RA, Veltkamp E, Nijkamp HJJ (1978) Isolation and characterization of a Clo DF13::Tn901 plasmid mutant with thermosensitive controle of DNA replication. J Bacteriol 135:612–621
Austin S, Ziese M, Sternberg N (1981) A novel role for site-specific recombination in maintenance of bacterial replicons. Cell 25:729–736
Barbour SD, Nagaishi H, Templin A, Clark AJ (1970) Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec + revertants caused by indirect suppression of Rec - mutations. Proc Natl Acad Sci USA 67:128–135
Embden JDA van (1978) Translocation of an ampicillin resistance determinant within an R-factor aggregate in Salmonella panama. Ant. v Leeuwenhoek 44:203–218
Embden JDA van, Veltkamp E, Stuitje AR, Andreoli PM, Nijkamp HJJ (1978) Integration of a transposable DNA sequence which mediates ampicillin resistance into Clo DF13 plasmid DNA: Determination of the site and orientation of Tn901 insertions. Plasmid 1:204–217
Fishel RA, James AA, Kolodner R (1981) RecA-independent general genetic recombination of plasmids. Nature 294:184–186
Gillen JR, Willis DK, Clark AJ (1981) Genetic analysis of the RecA pathway of genetic recombination in Escherichia coli K-12. J Bacteriol 145:521–532
Hakkaart MJJ, Veltkamp E, Nijkamp HJJ (1981) Protein H encoded by plasmid Clo DF13 involved in lysis of the bacterial host. I. Localisation of the gene and identification and subcellular localisation of the gene H product. Mol Gen Genet 183:318–325
Hakkaart MJJ, Wesseling JG, Veltkamp E, Nijkamp HJJ (1982) Maintenance of the bacteriocinogenic plasmid Clo DF13 in Escherichia coli cells. I. Localisation and mutual interactions of four Clo DF13 incompatibility regions. Mol Gen Genet 186:531–539
Heffron F, McCarthy BJ, Ohtsubo H, Ohtsubo E (1979) DNA sequence analysis of the transposon Tn3: three genes and three sites involved in transposition of Tn3 Cell 18:1153–1163
Laban A, Cohen A (1981) Interplasmidic and intraplasmidic recombination in Escherichia coli K-12. Mol Gen Genet 184:200–207
Nordström K, Molin S, Aagaard-Hansen H (1980) Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid 4:215–227
Pol H van de, Veltkamp E, Nijkamp HJJ (1980) Genetic information of the bacteriocinogenic plasmid Clo DF13 involved in the inhibition of the multiplication of double stranded DNA phages. Mol Gen Genet 178:535–540
Pol H van de (1980) The bacteriocinogenic plasmid Clo DF13: Mobilization and phage interaction. Dissertation Amsterdam Vrije Universiteit
Reed RR and Grindley NDF (1981) Transposon-mediated sitespecific recombination in vitro: DNA cleavage and protein —DNA linkage at the recombination site. Cell 25:721–728
Stuije AR, Veltkamp E, Elzen PJM van den, Nijkamp HJJ (1978) In vitro construction of deletion mutants of the bacteriocinogenic plasmid Clo DF13. Nucl Acids Res 5:1801–1820
Stuitje AR, Veltkamp E, Maat J, Heyneker HL (1980) The nucleotide sequence surrounding the replication origin of the cop3 mutant of the bacteriocinogenic plasmid Clo DF13. Nucl Acids Res 8:1459–1473
Stuitje AR, Spelt CE, Veltkamp E, Nijkamp HJJ (1981) Identification of mutations affecting replication control of plasmid Clo DF 13. Nature 290:264–267
Taylor A, Smith GR (1980) Unwinding and rewinding of DNA by the RecBC enzyme. Cell 22:447–457
Veltkamp E, Stuitje AR (1981) Replication and structure of the bacteriocinogenic plasmids Clo DF13 and Col E1. Plasmid 5:76–99
Veltkamp E, Barendsen W, Nijkamp HJJ (1974) Influence of protein and ribonucleic acid synthesis on the replication of the bacteriocinogenic factor Clo DF13 in Escherichia coli cells and minicells. J Bacteriol 118:165–174
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Communicated by H. Böhme
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Hakkaart, M.J.J., Veltkamp, E. & Niikamp, H.J.J. Maintenance of the bacteriocinogenic plasmid Clo DF13 in Escherichia coli cells. Mol Gen Genet 188, 338–344 (1982). https://doi.org/10.1007/BF00332698
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DOI: https://doi.org/10.1007/BF00332698