Summary
The nucleotide sequence of the cis-acting partition site (centromere) of the miniF plasmid has been determined. Its most notable feature is a reiterated 43 base pair unit. A series of plasmids deleted for portions of the repeat region was constructed and tested for incompatibility with R386 and for stability of inheritance. The extent of incompatibility with R386 was correlated with the number of repeat units. In contrast, the great majority of the repeats were not needed for miniF stability. An adjacent region of unique sequence was also found to be involved in centromere function.
Similar content being viewed by others
References
Austin S, Abeles A (1983) Partition of unit-copy miniplasmids to daughter cells II. The partition region of miniplasmid P1 encodes an essential protein and a centromere-like site at which it acts. J Mol Biol 169:373–387
Austin S, Wierzbecki A (1983) Two miniF-encoded proteins are essential for equipartitioning. Plasmid 10:73–81
Bergquist PL, Adelberg EA (1972) Abnormal excision and transfer of chromosomal segments by a strain of Escherichia coli K12. J Bacteriol 111:119–128
Bergquist PL, Downward RA, Caughey PA, Gardner RC, Lane HED (1981) Analysis of mini-F plasmid replication by transposition mutagenesis. J Bacteriol 147:888–899
Brandenburger A, Bailone A, Lévine A, Devoret R (1984) Gratuitous induction. J Mol Biol 179:571–576
Chattoraj DK, Abeles AL, Yarmolinsky MB (1985) P1 plasmid maintenance: a paradigm of precise control. In: Helinski DR, Cohen SN, Clewell D, Jackson DA, Hollaender A (eds) Plasmids in bacteria. Plenum Press, New York, pp 355–381
Eichenlaub R, Figurski D, Helinski DR (1977) Bidirectional replication from a unique origin in a mini-F plasmid. Proc Natl Acad Sci USA 74:1138–1141
Friedman S, Martin K, Austin S (1986) The partition system of the Pi plasmid. In: Evolution and environmental spread of antibiotic resistance genes. Cold Spring Harbour Laboratory, Cold Spring Harbour, New York (in press)
Gardner RC, Caughey PA, Lane D, Berquist PL (1980) Replication mutants of the F plasmid of Escherichia coli K12. II. Cloned replication regions of temperature-sensitive mutants. Plasmid 3:179–192
Gardner RC, Malcolm L, Bergquist PL, Lane HED (1982) IncD, a genetic locus in F responsible for incompatibility with several plasmids of the IncFI group. Mol Gen Genet 188:345–352
Hayakawa Y, Murotsu T, Matsubara K (1985) Mini-F protein that binds to a unique region for partition of mini-F plasmid DNA. J Bacteriol 163:349–354
Helsberg M, Eichenlaub R (1986) Twelve 43-base-pair repeats map in a cis-acting region essential for partition of plasmid mini-F. J Bacteriol 165:1043–1045
Hiraga S, Jaffé A, Ogura T, Mori H, Takahashi H (1986) F plasmid ccd mechanism in Escherichia coli. J Bacteriol 166:100–104
Jaffé A, Ogura T, Hiraga S (1985) Effects of ccd function of the F plasmid on bacterial growth. J Bacteriol 163:841–849
Jacob F, Brenner S, Cuzin F (1963) On the regulation of DNA replication in bacteria. Cold Spring Harbour Symp Quant Biol 28:329–348
Kahn ML, Figurski D, Ito L, Helinski DR (1979) Essential regions for replication of a stringent and a relaxed plasmid in Escherichia coli. Cold Spring Harbor Symp Quant Biol 43:99–103
Kline BC (1979) Incompatibility between F'lac, R386 and F:pSC101 recombinant plasmid: the specificity of F incompatibility genes. Plasmid 2:437–445
Lane D, Gardner RC (1979) Second EcoRI fragment of F capable of selfreplication. J Bacteriol 139:141–151
Lovett MA, Helinski DR (1976) Method for the isolation of the replication region of bacterial replicon: construction of a mini-F'Km plasmid. J Bacteriol 127:982–989
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manua. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, p 545
Manis JJ, Kline BC (1977) Restriction endonuclease mapping and mutagenesis of the F sex factor replication region. Mol Gen Genet 152:175–182
Maxam AM, Gilbert W (1977) A new method for sequencing DNA. Proc Natl Acad Sci USA 74:560–564
Meyers JA, Sanchez D, Elwell LP, Falkow S (1976) Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol 127:1529–1537
Nordström K, Molin S, Aagaard-Hansen H (1980) Partitioning of plasmid R1 in E. coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid 4:215–227
Ogura T, Hiraga S (1983) Partition mechanism of F plasmid: two plasmid gene-encoded products and a cis-acting region are involved in partition. Cell 32:351–360
Ogura T, Miki T, Hiraga S (1980) Copy-number mutants of the plasmid carryiing the replication origin of the Escherichia coli chromosome: evidence for a control region of replication. Proc Natl Acad Sci USA 77:3993–3997
Palchaudhuri S, Maas WK (1977) Physical mapping of a DNA sequence common to plasmids of incompatibility group FI. Proc Natl Acad Sci USA 74:1190–1194
Phua S-H, Bergquist PL, Lane HED (1982) Effects of Tn5 insertion in the incD region on mini-F maintenance and polypeptide synthesis. Mol Gen Genet 188:353–355
Timmis KN, Cabello F, Cohen SN (1975) Cloning, isolation and characterization of replication regions of complex plasmid genomes. Proc Natl Acad Sci USA 72:2242–2246
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Watson LA, Phus S-H, Bergquist PL, Lane HED (1982) An Mr29000 protein is essential for mini-F maintenance in E. coli. Gene 19:173–178
Author information
Authors and Affiliations
Additional information
Communicated by R. Devoret
Rights and permissions
About this article
Cite this article
Lane, D., Rothenbuehler, R., Merrillat, AM. et al. Analysis of the F plasmid centromere. Mol Gen Genet 207, 406–412 (1987). https://doi.org/10.1007/BF00331608
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331608