Skip to main content
NCBI home page
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
. 1985 Jun;82(11):3776–3780. doi: 10.1073/pnas.82.11.3776

Site-specific DNA inversion is enhanced by a DNA sequence element in cis

Hans E Huber 1,*, Shigeru Iida 1,†, Werner Arber 1, Thomas A Bickle 1
PMCID: PMC397870  PMID: 16593573

Abstract

A segment of the bacteriophage P1 genome, called the C segment, can be inverted by site-specific recombination; the two different orientations of the invertible segment confer different host ranges to the phage. Inversion is catalyzed by the product of the cin gene which is adjacent to one of the crossover sites flanking the C segment. The Cin-catalyzed recombination can be measured in trans by using tester plasmids in which inversion switches on antibiotic-resistance genes. We show here that an additional sequence, distinct from the two crossover sites, is needed in cis for efficient inversion. This sequence is part of the cin structural gene and stimulates recombination more than 100-fold. We have localized the major enhancer sequence on a 72-base-pair fragment and found its activity to be largely independent of the orientation or position of the sequence with respect to the crossover sites.

Keywords: site-specific recombination, recombinational enhancer, BAL-31 deletion analysis

Full text

PDF
3776

Images in this article

3778Image
on p.3778
3778Image
on p.3778
3778Image
on p.3778
3778Image
on p.3778
3778Image
on p.3778

Selected References

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

  1. Giphart-Gassler M., Plasterk R. H., van de Putte P. G inversion in bacteriophage Mu: a novel way of gene splicing. Nature. 1982 May 27;297(5864):339–342. doi: 10.1038/297339a0. [DOI] [PubMed] [Google Scholar]
  2. Hiestand-Nauer R., Iida S. Sequence of the site-specific recombinase gene cin and of its substrates serving in the inversion of the C segment of bacteriophage P1. EMBO J. 1983;2(10):1733–1740. doi: 10.1002/j.1460-2075.1983.tb01650.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Huber H. E., Iida S., Bickle T. A. Expression of the bacteriophage P1 cin recombinase gene from its own and heterologous promoters. Gene. 1985;34(1):63–72. doi: 10.1016/0378-1119(85)90295-1. [DOI] [PubMed] [Google Scholar]
  4. Iida S. Bacteriophage P1 carries two related sets of genes determining its host range in the invertible C segment of its genome. Virology. 1984 Apr 30;134(2):421–434. doi: 10.1016/0042-6822(84)90309-x. [DOI] [PubMed] [Google Scholar]
  5. Iida S., Huber H., Hiestand-Nauer R., Meyer J., Bickle T. A., Arber W. The bacteriophage P1 site-specific recombinase cin: recombination events and DNA recognition sequences. Cold Spring Harb Symp Quant Biol. 1984;49:769–777. doi: 10.1101/sqb.1984.049.01.087. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Iino T., Kutsukake K. Trans-acting genes of bacteriophages P1 and Mu mediate inversion of a specific DNA segment involved in flagellar phase variation of Salmonella. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):11–16. doi: 10.1101/sqb.1981.045.01.004. [DOI] [PubMed] [Google Scholar]
  8. Ikemura T. Correlation between the abundance of Escherichia coli transfer RNAs and the occurrence of the respective codons in its protein genes: a proposal for a synonymous codon choice that is optimal for the E. coli translational system. J Mol Biol. 1981 Sep 25;151(3):389–409. doi: 10.1016/0022-2836(81)90003-6. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Kahmann R., Rudt F., Mertens G. Substrate and enzyme requirements for in vitro site-specific recombination in bacteriophage mu. Cold Spring Harb Symp Quant Biol. 1984;49:285–294. doi: 10.1101/sqb.1984.049.01.034. [DOI] [PubMed] [Google Scholar]
  11. Kamp D., Chow L. T., Broker T. R., Kwoh D., Zipser D., Kahmann R. Site-specific recombination in phage mu. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1159–1167. doi: 10.1101/sqb.1979.043.01.131. [DOI] [PubMed] [Google Scholar]
  12. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  13. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mertens G., Hoffmann A., Blöcker H., Frank R., Kahmann R. Gin-mediated site-specific recombination in bacteriophage Mu DNA: overproduction of the protein and inversion in vitro. EMBO J. 1984 Oct;3(10):2415–2421. doi: 10.1002/j.1460-2075.1984.tb02148.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nash H. A., Mizuuchi K., Enquist L. W., Weisberg R. A. Strand exchange in lambda integrative recombination: genetics, biochemistry, and models. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):417–428. doi: 10.1101/sqb.1981.045.01.056. [DOI] [PubMed] [Google Scholar]
  16. Plasterk R. H., Brinkman A., van de Putte P. DNA inversions in the chromosome of Escherichia coli and in bacteriophage Mu: relationship to other site-specific recombination systems. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5355–5358. doi: 10.1073/pnas.80.17.5355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Plasterk R. H., Kanaar R., van de Putte P. A genetic switch in vitro: DNA inversion by Gin protein of phage Mu. Proc Natl Acad Sci U S A. 1984 May;81(9):2689–2692. doi: 10.1073/pnas.81.9.2689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Schmitz A. Cyclic AMP receptor proteins interacts with lactose operator DNA. Nucleic Acids Res. 1981 Jan 24;9(2):277–292. doi: 10.1093/nar/9.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Silverman M., Zieg J., Mandel G., Simon M. Analysis of the functional components of the phase variation system. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):17–26. doi: 10.1101/sqb.1981.045.01.005. [DOI] [PubMed] [Google Scholar]
  23. Stahl F. W. Special sites in generalized recombination. Annu Rev Genet. 1979;13:7–24. doi: 10.1146/annurev.ge.13.120179.000255. [DOI] [PubMed] [Google Scholar]
  24. Sternberg N., Hoess R. The molecular genetics of bacteriophage P1. Annu Rev Genet. 1983;17:123–154. doi: 10.1146/annurev.ge.17.120183.001011. [DOI] [PubMed] [Google Scholar]
  25. Wells R. G., Grindley N. D. Analysis of the gamma delta res site. Sites required for site-specific recombination and gene expression. J Mol Biol. 1984 Nov 15;179(4):667–687. doi: 10.1016/0022-2836(84)90161-x. [DOI] [PubMed] [Google Scholar]
  26. Zieg J., Simon M. Analysis of the nucleotide sequence of an invertible controlling element. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4196–4200. doi: 10.1073/pnas.77.7.4196. [DOI] [PMC free article] [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