Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Feb;87(3):1109–1113. doi: 10.1073/pnas.87.3.1109

Site-specific DNA recombination system Min of plasmid p15B: a cluster of overlapping invertible DNA segments.

H Sandmeier 1, S Iida 1, J Meyer 1, R Hiestand-Nauer 1, W Arber 1
PMCID: PMC53420  PMID: 2405394

Abstract

Plasmid p15B of Escherichia coli 15T- carries a 3.5-kilobase segment that undergoes frequent DNA inversion mediated by the DNA inversion enzyme Min, a member of the Din family of site-specific recombinases. While the previously described Din inversion systems invert a DNA segment between two crossover sites in inverted orientation, the Min system produces more complex DNA rearrangements. These have been physically characterized by electron microscopy and by restriction cleavage analysis. The results can best be explained by a model that involves six crossover sites (called mix) and predicts 240 isomeric forms of the invertible region. The model was confirmed by sequencing the six mix sites in plasmids that contain the invertible DNA segments in a frozen configuration. All mix sites fit the dix consensus sequence, and they are all good substrates for DNA inversion when carried in inverted orientation. Recombination between two mix sites in direct orientation was rare, in line with the notion that Din inversion systems are topologically biased to the inversion reaction. Another recently described multiple inversion system, the shufflon of the E. coli plasmid R64, is neither functionally nor structurally related to the Min system of p15B.

Full text

PDF
1109

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arber W., Wauters-Willems D. Host specificity of DNA produced by Escherichia coli. XII. The two restriction and modification systems of strain 15T-. Mol Gen Genet. 1970;108(3):203–217. doi: 10.1007/BF00283350. [DOI] [PubMed] [Google Scholar]
  2. Austin S., Ziese M., Sternberg N. A novel role for site-specific recombination in maintenance of bacterial replicons. Cell. 1981 Sep;25(3):729–736. doi: 10.1016/0092-8674(81)90180-x. [DOI] [PubMed] [Google Scholar]
  3. Craig N. L. The mechanism of conservative site-specific recombination. Annu Rev Genet. 1988;22:77–105. doi: 10.1146/annurev.ge.22.120188.000453. [DOI] [PubMed] [Google Scholar]
  4. Haffter P., Bickle T. A. Enhancer-independent mutants of the Cin recombinase have a relaxed topological specificity. EMBO J. 1988 Dec 1;7(12):3991–3996. doi: 10.1002/j.1460-2075.1988.tb03287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Iida S., Hiestand-Nauer R. Localized conversion at the crossover sequences in the site-specific DNA inversion system of bacteriophage P1. Cell. 1986 Apr 11;45(1):71–79. doi: 10.1016/0092-8674(86)90539-8. [DOI] [PubMed] [Google Scholar]
  6. Iida S., Hiestand-Nauer R. Role of the central dinucleotide at the crossover sites for the selection of quasi sites in DNA inversion mediated by the site-specific Cin recombinase of phage P1. Mol Gen Genet. 1987 Jul;208(3):464–468. doi: 10.1007/BF00328140. [DOI] [PubMed] [Google Scholar]
  7. Iida S., Meyer J., Kennedy K. E., Arber W. A site-specific, conservative recombination system carried by bacteriophage P1. Mapping the recombinase gene cin and the cross-over sites cix for the inversion of the C segment. EMBO J. 1982;1(11):1445–1453. doi: 10.1002/j.1460-2075.1982.tb01336.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Johnson R. C., Bruist M. F. Intermediates in Hin-mediated DNA inversion: a role for Fis and the recombinational enhancer in the strand exchange reaction. EMBO J. 1989 May;8(5):1581–1590. doi: 10.1002/j.1460-2075.1989.tb03542.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kanaar R., van de Putte P., Cozzarelli N. R. Gin-mediated recombination of catenated and knotted DNA substrates: implications for the mechanism of interaction between cis-acting sites. Cell. 1989 Jul 14;58(1):147–159. doi: 10.1016/0092-8674(89)90411-x. [DOI] [PubMed] [Google Scholar]
  10. Kennedy K. E., Iida S., Meyer J., Stålhammar-Carlemalm M., Hiestand-Nauer R., Arber W. Genome fusion mediated by the site specific DNA inversion system of bacteriophage P1. Mol Gen Genet. 1983;189(3):413–421. doi: 10.1007/BF00325903. [DOI] [PubMed] [Google Scholar]
  11. Klippel A., Mertens G., Patschinsky T., Kahmann R. The DNA invertase Gin of phage Mu: formation of a covalent complex with DNA via a phosphoserine at amino acid position 9. EMBO J. 1988 Apr;7(4):1229–1237. doi: 10.1002/j.1460-2075.1988.tb02935.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Komano T., Kubo A., Kayanuma T., Furuichi T., Nisioka T. Highly mobile DNA segment of IncI alpha plasmid R64: a clustered inversion region. J Bacteriol. 1986 Jan;165(1):94–100. doi: 10.1128/jb.165.1.94-100.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Komano T., Kubo A., Nisioka T. Shufflon: multi-inversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames. Nucleic Acids Res. 1987 Feb 11;15(3):1165–1172. doi: 10.1093/nar/15.3.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kubo A., Kusukawa A., Komano T. Nucleotide sequence of the rci gene encoding shufflon-specific DNA recombinase in the IncI1 plasmid R64: homology to the site-specific recombinases of integrase family. Mol Gen Genet. 1988 Jul;213(1):30–35. doi: 10.1007/BF00333394. [DOI] [PubMed] [Google Scholar]
  15. Meyer J., Stålhammar-Carlemalm M., Streiff M., Iida S., Arber W. Sequence relations among the IncY plasmid p15B, P1, and P7 prophages. Plasmid. 1986 Sep;16(2):81–89. doi: 10.1016/0147-619x(86)90066-1. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Scott T. N., Simon M. I. Genetic analysis of the mechanism of the Salmonella phase variation site specific recombination system. Mol Gen Genet. 1982;188(2):313–321. doi: 10.1007/BF00332694. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES