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. 1988 Jul;85(13):4628–4632. doi: 10.1073/pnas.85.13.4628

FLP recombinase is an enzyme.

C A Gates 1, M M Cox 1
PMCID: PMC280488  PMID: 3290899

Abstract

The FLP protein of the yeast 2-microns plasmid catalyzes intermolecular site-specific recombination with a turnover number of approximately equal to 0.12 min-1 (per FLP monomer) for relaxed DNA substrates. Under conditions that enhance its stability, the protein can be used in catalytic rather than stoichiometric amounts. The reaction rate exhibits a strong dependence on FLP protein concentration even when the protein is present in excess relative to available recombination sites.

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Selected References

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  1. Abremski K., Hoess R. Bacteriophage P1 site-specific recombination. Purification and properties of the Cre recombinase protein. J Biol Chem. 1984 Feb 10;259(3):1509–1514. [PubMed] [Google Scholar]
  2. Abremski K., Hoess R., Sternberg N. Studies on the properties of P1 site-specific recombination: evidence for topologically unlinked products following recombination. Cell. 1983 Apr;32(4):1301–1311. doi: 10.1016/0092-8674(83)90311-2. [DOI] [PubMed] [Google Scholar]
  3. Andrews B. J., McLeod M., Broach J., Sadowski P. D. Interaction of the FLP recombinase of the Saccharomyces cerevisiae 2 micron plasmid with mutated target sequences. Mol Cell Biol. 1986 Jul;6(7):2482–2489. doi: 10.1128/mcb.6.7.2482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andrews B. J., Proteau G. A., Beatty L. G., Sadowski P. D. The FLP recombinase of the 2 micron circle DNA of yeast: interaction with its target sequences. Cell. 1985 Apr;40(4):795–803. doi: 10.1016/0092-8674(85)90339-3. [DOI] [PubMed] [Google Scholar]
  5. Bruckner R. C., Cox M. M. Specific contacts between the FLP protein of the yeast 2-micron plasmid and its recombination site. J Biol Chem. 1986 Sep 5;261(25):11798–11807. [PubMed] [Google Scholar]
  6. Cox M. M. The FLP protein of the yeast 2-microns plasmid: expression of a eukaryotic genetic recombination system in Escherichia coli. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4223–4227. doi: 10.1073/pnas.80.14.4223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Futcher A. B. Copy number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae. J Theor Biol. 1986 Mar 21;119(2):197–204. doi: 10.1016/s0022-5193(86)80074-1. [DOI] [PubMed] [Google Scholar]
  8. Gellert M., Nash H. Communication between segments of DNA during site-specific recombination. 1987 Jan 29-Feb 4Nature. 325(6103):401–404. doi: 10.1038/325401a0. [DOI] [PubMed] [Google Scholar]
  9. Gronostajski R. M., Sadowski P. D. The FLP protein of the 2-micron plasmid of yeast. Inter- and intramolecular reactions. J Biol Chem. 1985 Oct 5;260(22):12328–12335. [PubMed] [Google Scholar]
  10. Gronostajski R. M., Sadowski P. D. The FLP recombinase of the Saccharomyces cerevisiae 2 microns plasmid attaches covalently to DNA via a phosphotyrosyl linkage. Mol Cell Biol. 1985 Nov;5(11):3274–3279. doi: 10.1128/mcb.5.11.3274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hatfull G. F., Noble S. M., Grindley N. D. The gamma delta resolvase induces an unusual DNA structure at the recombinational crossover point. Cell. 1987 Apr 10;49(1):103–110. doi: 10.1016/0092-8674(87)90760-4. [DOI] [PubMed] [Google Scholar]
  12. Johnson R. C., Bruist M. B., Glaccum M. B., Simon M. I. In vitro analysis of Hin-mediated site-specific recombination. Cold Spring Harb Symp Quant Biol. 1984;49:751–760. doi: 10.1101/sqb.1984.049.01.085. [DOI] [PubMed] [Google Scholar]
  13. Kostriken R., Morita C., Heffron F. Transposon Tn3 encodes a site-specific recombination system: identification of essential sequences, genes, and actual site of recombination. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4041–4045. doi: 10.1073/pnas.78.7.4041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McLeod M., Craft S., Broach J. R. Identification of the crossover site during FLP-mediated recombination in the Saccharomyces cerevisiae plasmid 2 microns circle. Mol Cell Biol. 1986 Oct;6(10):3357–3367. doi: 10.1128/mcb.6.10.3357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meyer-Leon L., Gates C. A., Attwood J. M., Wood E. A., Cox M. M. Purification of the FLP site-specific recombinase by affinity chromatography and re-examination of basic properties of the system. Nucleic Acids Res. 1987 Aug 25;15(16):6469–6488. doi: 10.1093/nar/15.16.6469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Meyer-Leon L., Senecoff J. F., Bruckner R. C., Cox M. M. Site-specific genetic recombination promoted by the FLP protein of the yeast 2-micron plasmid in vitro. Cold Spring Harb Symp Quant Biol. 1984;49:797–804. doi: 10.1101/sqb.1984.049.01.090. [DOI] [PubMed] [Google Scholar]
  17. Nash H. A., Bauer C. E., Gardner J. F. Role of homology in site-specific recombination of bacteriophage lambda: evidence against joining of cohesive ends. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4049–4053. doi: 10.1073/pnas.84.12.4049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nash H. A. Integration and excision of bacteriophage lambda: the mechanism of conservation site specific recombination. Annu Rev Genet. 1981;15:143–167. doi: 10.1146/annurev.ge.15.120181.001043. [DOI] [PubMed] [Google Scholar]
  19. Nash H. A., Robertson C. A. Purification and properties of the Escherichia coli protein factor required for lambda integrative recombination. J Biol Chem. 1981 Sep 10;256(17):9246–9253. [PubMed] [Google Scholar]
  20. Nunes-Düby S. E., Matsumoto L., Landy A. Site-specific recombination intermediates trapped with suicide substrates. Cell. 1987 Aug 28;50(5):779–788. doi: 10.1016/0092-8674(87)90336-9. [DOI] [PubMed] [Google Scholar]
  21. Plasterk R. H., Ilmer T. A., Van de Putte P. Site-specific recombination by Gin of bacteriophage Mu: inversions and deletions. Virology. 1983 May;127(1):24–36. doi: 10.1016/0042-6822(83)90367-7. [DOI] [PubMed] [Google Scholar]
  22. Prasad P. V., Horensky D., Young L. J., Jayaram M. Substrate recognition by the 2 micron circle site-specific recombinase: effect of mutations within the symmetry elements of the minimal substrate. Mol Cell Biol. 1986 Dec;6(12):4329–4334. doi: 10.1128/mcb.6.12.4329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Reed R. R. Transposon-mediated site-specific recombination: a defined in vitro system. Cell. 1981 Sep;25(3):713–719. doi: 10.1016/0092-8674(81)90178-1. [DOI] [PubMed] [Google Scholar]
  24. Reynolds A. E., Murray A. W., Szostak J. W. Roles of the 2 microns gene products in stable maintenance of the 2 microns plasmid of Saccharomyces cerevisiae. Mol Cell Biol. 1987 Oct;7(10):3566–3573. doi: 10.1128/mcb.7.10.3566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sadowski P. Site-specific recombinases: changing partners and doing the twist. J Bacteriol. 1986 Feb;165(2):341–347. doi: 10.1128/jb.165.2.341-347.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Senecoff J. F., Bruckner R. C., Cox M. M. The FLP recombinase of the yeast 2-micron plasmid: characterization of its recombination site. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7270–7274. doi: 10.1073/pnas.82.21.7270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Senecoff J. F., Cox M. M. Directionality in FLP protein-promoted site-specific recombination is mediated by DNA-DNA pairing. J Biol Chem. 1986 Jun 5;261(16):7380–7386. [PubMed] [Google Scholar]
  28. Thompson J. F., Moitoso de Vargas L., Koch C., Kahmann R., Landy A. Cellular factors couple recombination with growth phase: characterization of a new component in the lambda site-specific recombination pathway. Cell. 1987 Sep 11;50(6):901–908. doi: 10.1016/0092-8674(87)90516-2. [DOI] [PubMed] [Google Scholar]
  29. Vetter D., Andrews B. J., Roberts-Beatty L., Sadowski P. D. Site-specific recombination of yeast 2-micron DNA in vitro. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7284–7288. doi: 10.1073/pnas.80.23.7284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Volkert F. C., Broach J. R. Site-specific recombination promotes plasmid amplification in yeast. Cell. 1986 Aug 15;46(4):541–550. doi: 10.1016/0092-8674(86)90879-2. [DOI] [PubMed] [Google Scholar]

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