Abstract
The yeast 2-micron plasmid is a 6,318-bp circular DNA molecule occurring in many yeast strains. It is generally present at 50 to 100 copies per cell and has been sequenced in its entirety (11). The plasmid encodes at least four protein products (Ref. 6; Volkert et al., this Volume), which are involved primarily in plasmid maintenance. One of these proteins, designated FLP, is the central component in a plasmid-encoded, site-specific recombination system. The sequence of the open reading frame (ORF) encoding this protein provides a predicted molecular weight of slightly less than 48,000 Da. This recombinase acts at a pair of sites located within the 599-bp inverted repeats. Recombination results in the inversion of the DNA between the sites, giving two forms of the plasmid, termed A and B (see Fig. 1). Isolated plasmid DNA is found to be a 50:50 mixture of the two forms. Inversion has been shown to be part of a copy-number amplification system (Volkert et al., this Volume). The general features of the plasmid and its maintenance systems appear to be shared by plasmids isolated from a variety of related organisms (A. Toh-e et al., this Volume).
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References
Andrews, B.J., M. McLeod, J. Broach, and P.D. Sadowski (1986) Interaction of the FLP recombinase of the yeast 2-micron plasmid with mutated target sequences. Mol. Cell. Biol. (in press).
Andrews, B.J., G.A. Proteau, L.G. Beatty, and P.D. Sadowski (1985) The FLP recombinase of the 2-micron circle DNA of yeast: Interaction with its target sequences. Cell 40:795–803.
Babineau, D., D. Vetter, B.J. Andrews, R.M. Gronostajski, G.A. Proteau, L.G. Beatty, and P.D. Sadowski (1985) The FLP protein of the 2-micron plasmid of yeast: Purification of the protein from Escherichia coli cells expressing the cloned FLP gene. J. Biol. Chem. 260:12313–12319.
Bauer, C.E., S.D. Hesse, J.F. Gardner, and R.I. Gumport (1984) DNA interactions during bacteriophage X site-specific recombination. Cold Spring Harbor Symp. Quant. Biol. 49:699–705.
Benjamin, H.W., M.M. Matzuk, M.A. Krasnow, and N.R. Cozzarelli (1985) Recombination site selection by Tn3 resolvase: Topological tests of a tracking mechanism. Cell 49:147–158.
Broach, J.R., V.R. Guarascio, and M. Jayaram (1982) Recombination with the yeast 2 micron plasmid is site-specific. Cell 29:227–234.
Bruckner, R.C., and M.M. Cox (1986) Specific contacts between the FLP protein of the yeast 2 micron plasmid and its recombination site. J. Biol. Chem. (in press).
Cox, M.M. (1983) The FLP protein of the yeast 2 micron plasmid: Expression of a eukaryotic genetic recombination system in Escherichia coli. Proc. Natl. Acad. Sci., USA 80:4223–4227.
Gronastajski, R.M., and P.D. Sadowski (1985) Determination of DNA sequences essential for FLP-mediated recombination by a novel method. J. Biol. Chem. 260:12320–12327.
Gronastajski, R.M., and P.D. Sadowski (1985) The FLP recombinase of the yeast 2-micron plasmid attaches covalently to DNA via a phosphotyrosyl linkage. Mol. Cell. Biol. 5:3274–3279.
Hartley, J.L., and J.E. Donelson (1980) Nucleotide sequence of the yeast plasmid. Nature (London) 268:860–864.
Hoess, R.H., A. Wierzbicki, and K.A. Abremski (1986) The role of the loxP spacer region in Pl site-specific recombination. Nucl. Acids Res. 14:2287–2300.
Jayaram, M. (1985) Two-micrometer circle site-specific recombination: The minimal substrate and the possible role of flanking sequences. Proc. Natl. Acad. Sci., USA 82:5875–5879.
Johnson, R.C., and M.I. Simon (1985) Hin-mediated site-specific recombination requires two 26 base pair recombination sites and a 60 base pair recombinational enhancer. Cell 41:784–791.
Kikuchi, Y., and H. Nash (1979) Nicking-closing activity associated with bacteriophage X int gene product. Proc. Natl. Acad. Sci., USA 76:3760–3764.
Kitts, P., E. Richet, and H. Nash (1984) X Integrative recombination; supercoiling, synapsis, and strand exchange. Cold Spring Harbor Symp. Quant. Biol. 49:735–744.
Meyer-Leon, L., J.F. Senecoff, R.C. Bruckner, and M.M. Cox (1984) Site-specific recombination promoted by the FLP protein of the yeast 2-micron plasmid in vitro. Cold Spring Harbor Symp. Quant. Biol. 49:797–804.
Reed, R.R., and N.D.F. Grindley (1981) Transposon-mediated site-specific recombination: A defined in vitro system. Cell 25:713–719.
Sadowski, P.D., D.D. Lee, B.J. Andrews, D. Babineau, L. Beatty, M.J. Morse, G.A. Proteau, and D. Vetter (1984) In vitro systems for the genetic recombination of the DNAs of bacteriophage T7 and yeast 2-micron circle. Cold Spring Harbor Symp. Quant. Biol. 49:789–796.
Senecoff, J.F., and M.M. Cox (1986) Directionality in FLP protein-promoted site-specific recombination is mediated by DNA-DNA pairing. J. Biol. Chem. 261:7380–7386.
Senecoff, J.F., R.C. Bruckner, and M.M. Cox (1985) The FLP recombinase of the yeast 2-micron plasmid: Characterization of its recombination site. Proc. Natl. Acad. Sci., USA 82:7270–7274.
Vetter, D., B.J. Andrews, L. Roberts-Beatty, and P.D. Sadowski (1983) Site-specific recombination of the yeast 2 micron DNA in vitro. Proc. Natl. Acad. Sci., USA 80:7284–7288.
Weisberg, R.A., L.W. Enquist, C. Foeller, and A. Landy (1983) Role for DNA homology in site-specific recombination. The isolation and characterization of a site affinity mutant of coliphage λ. J. Mol. Biol. 170:318–342.
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© 1986 Plenum Press, New York
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Senecoff, J.F. et al. (1986). Site-Specific Recombination Promoted in Vitro by the FLP Protein of the Yeast Two-Micron Plasmid. In: Wickner, R.B., et al. Extrachromosomal Elements in Lower Eukaryotes. Basic Life Sciences, vol 40. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5251-8_30
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DOI: https://doi.org/10.1007/978-1-4684-5251-8_30
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