Abstract
We have studied the recombination of plasmids bearing bom and cer sites. The bom (basis of mobilization) site is required for conjugative transfer, while the cer (ColE1 resolution) site is involved in the resolution of plasmid multimers, which increases plasmid stability. We constructed a pair of parent plasmids in such a way as to allow us select clones containing recombinant plasmids directly. Clone selection was based on the McrA sensitivity of recipient host DNA modified by M. Ecl18kI, which is encoded by one of the parent plasmids. The recombinant plasmid contains segments originating from both parental DNAs, which are bounded by bom and cer sites. Its structure is in accordance with our previously proposed model for recombination mediated by bom and cer sequences. The frequency of recombinant plasmid formation coincided with the frequency of recombination at the bom site. We also show that bom-mediated recombination in trans, unlike in cis, is independent of other genetic determinants on the conjugative plasmids.
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References
Albiger B, Comanducci A, Dodd HM, Tavakoli N, Lett MC, Bennett P (2000) pUB2380: characterization of a ColD-like resistance plasmid. Plasmid 44:54–65
Bastia D (1978) Determination of restriction sites and the nucleotide sequence surrounding the relaxation site of ColE1. J Mol Biol 124:601–639
Boyd AC, Archer JA, Sherratt DJ (1989) Characterization of the ColE1 mobilization region and its protein products. Mol Gen Genet 217:488–498
Chan PT, Ohmori H, Tomizawa J, Lebowitz J (1985) Nucleotide sequence and gene organization of ColE1 DNA. J Biol Chem 260:8925–8935.
Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in Escherichia coli: Purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166
Denjmukhametov MM, Zakharova MV, Kravets AN, Pertsev AV, Sineva EV, Repik AV, Beletskaya IV, Gromova ES, Solonin AS (1997) Characterization of plasmids encoding type II restriction-modification systems, isoschizomers of SsoII. Mol Biol (Mosk) 31:708–714
Hashimoto-Gotoh T, Franklin FC, Nordheim A, Timmis KN (1981) Specific-purpose plasmid cloning vectors. I. Low copy number, temperature-sensitive, mobilization-defective pSC101-derived containment vectors. Gene 16:227–35
Ibanes M, Alvares I, Rodriguez-Pena JM, Rotger R (1996) A ColEI-type plasmid from Salmonella enteritidis encodes a DNA cytosine methyltransferase. Gene 196:145–158
Rehel N, Szatmari G (1996) Characterization of the stable maintenance of the Shigella flexneri plasmid pHS-2. Plasmid 36:183–190
Keenleyside WJ, Whitfield C (1995) Lateral transfer of rfb genes: a mobilizable ColE1-type plasmid carries the rfb O:54 (O:54 antigen biosynthesis) gene cluster from Salmonella enterica serovar Borreze. J Bacteriol 177:5247–5253
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual (2nd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Stirling CJ, Szatmari G, Stewart G, Smith MC, Sherratt DJ (1988) The arginine repressor is essential for plasmid-stabilizing site-specific recombination at the ColE1 cer locus. EMBO J 7:4389–95.
Subramanya HS, Arciszewska LK, Baker RA, Bird LE, Sherratt DJ, Wigley DB (1997) Crystal structure of the site-specific recombinase, XerD. EMBO J 16:5178–5187
Summers DK (1998) Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability. Mol Microbiol 29:1137–1145
Tomizawa JI, Ohmori H, Bird RE (1977) Origin of replication of colicin E1 plasmid DNA. Proc Natl Acad Sci USA 74:1865–1869
Warren GJ, Clark AJ (1980) Sequence-specific recombination of plasmid ColE1. Proc Natl Acad Sci USA 77:6724–6728
Zaitzev EN, Zaitzeva EM, Bakhlamova IV, Gorelov VN, Kuzmin NP, Kryukov VM, Lanzov VA (1986) Cloning and characterization of rec A gene from Pseudomonas aeruginosa (in Russian). Genetika 22:2721–2727
Zakharova MV, Pertzev AV, Kravetz AN, Beletskaya IV, Shlyapnikov MG, Solonin AS (1998) Complete nucleotide sequence of the Hsd plasmid pECO29 and identification of its functional regions. Biochim Biophys Acta 1398:106–112
Zakharova MV, Beletskaya IV, Denjmukhametov MM, Yurkova TV, Semenova LM, Shlyapnikov MG, Solonin AS (2002) Characterization of pECL18 and pKPN2: a proposed pathway for the evolution of two plasmids that carry identical genes for a Type II restriction-modification system. Mol Genet Genomics 267:171–178
Zakova N, Szatmari GB (1995) Site-specific recombination between ColE1 cer and NTP16 nmr sites in vivo. Mol Gen Genet 247:509–514
Acknowledgements
The authors thank Dr. A. A. Zimin and S. Mayorov for fruitful discussions and Dr. T. V. Ivashina for providing the F(lac-gal), tetr plasmid. Corrections to the English version of the manuscript made by E. Serebrova and V. Mikoyan are gratefully acknowledged
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Zakharova, M.V., Beletskaya, I.V., Bolovin, D.V. et al. Structural plasmid evolution as a result of coupled recombinations at bom and cer sites. Mol Genet Genomics 270, 415–419 (2004). https://doi.org/10.1007/s00438-003-0933-0
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DOI: https://doi.org/10.1007/s00438-003-0933-0