Skip to main content
PMC 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
. 1986 Aug;83(15):5474–5478. doi: 10.1073/pnas.83.15.5474

Integration of staphylococcal phage L54a occurs by site-specific recombination: structural analysis of the attachment sites.

C Y Lee, J J Iandolo
PMCID: PMC386309  PMID: 2942938

Abstract

Lysogenization by staphylococcal phage L54a induces the loss of lipase (glycerol ester hydrolase) activity in its host Staphylococcus aureus. The attachment site of the bacterial chromosome (attB) for the phage is at the 3' end of the lipase gene, geh. The DNA fragment containing the attB (base pairs 2620-2637 inclusive) site has been sequenced. We have also cloned and determined the nucleotide sequence of the DNA fragments containing the other three attachment sites--i.e., the attP locus on the circularly permuted phage genome and the attL and attR loci at the left and right ends of the prophage in the lysogenized strain. These results reveal that an 18-base-pair core sequence is common to all four att sites. These data indicate that the crossover point must exist within the core sequence and, further, that integration is site- and orientation-specific. We also localized the viral recombinase gene to a 2.1-kilobase DNA segment extending rightward to the attP site. This region was found to be essential for integration of plasmids containing the attP site.

Full text

PDF
5475

Images in this article

5477Image
on p.5477

Selected References

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

  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. Botchan P. An electron microscopic comparison of transcription on linear and superhelical DNA. J Mol Biol. 1976 Jul 25;105(1):161–176. doi: 10.1016/0022-2836(76)90201-1. [DOI] [PubMed] [Google Scholar]
  3. Brack C., Bickle T. A., Yuan R. The relation of single-stranded regions in bacteriophage PM2 supercoiled DNA to the early melting sequences. J Mol Biol. 1975 Aug 25;96(4):693–702. doi: 10.1016/0022-2836(75)90146-1. [DOI] [PubMed] [Google Scholar]
  4. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  5. Craig N. L., Nash H. A. E. coli integration host factor binds to specific sites in DNA. Cell. 1984 Dec;39(3 Pt 2):707–716. doi: 10.1016/0092-8674(84)90478-1. [DOI] [PubMed] [Google Scholar]
  6. Davies R. W., Schreier P. H., Kotewicz M. L., Echols H. Studies on the binding of lambda Int protein to attachment site DNA: identification of a tight-binding site in the P' region. Nucleic Acids Res. 1979 Dec 20;7(8):2255–2273. doi: 10.1093/nar/7.8.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duval-Iflah Y. Lysogenic conversion of the lipase in Staphylococcus pyogenes group 3 strains. Can J Microbiol. 1972 Sep;18(9):1491–1497. doi: 10.1139/m72-228. [DOI] [PubMed] [Google Scholar]
  8. Hoess R. H., Foeller C., Bidwell K., Landy A. Site-specific recombination functions of bacteriophage lambda: DNA sequence of regulatory regions and overlapping structural genes for Int and Xis. Proc Natl Acad Sci U S A. 1980 May;77(5):2482–2486. doi: 10.1073/pnas.77.5.2482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hoess R. H., Ziese M., Sternberg N. P1 site-specific recombination: nucleotide sequence of the recombining sites. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3398–3402. doi: 10.1073/pnas.79.11.3398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hsu P. L., Ross W., Landy A. The lambda phage att site: functional limits and interaction with Int protein. Nature. 1980 May 8;285(5760):85–91. doi: 10.1038/285085a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Johnson R. C., Simon M. I. Hin-mediated site-specific recombination requires two 26 bp recombination sites and a 60 bp recombinational enhancer. Cell. 1985 Jul;41(3):781–791. doi: 10.1016/s0092-8674(85)80059-3. [DOI] [PubMed] [Google Scholar]
  12. Kahmann R., Rudt F., Koch C., Mertens G. G inversion in bacteriophage Mu DNA is stimulated by a site within the invertase gene and a host factor. Cell. 1985 Jul;41(3):771–780. doi: 10.1016/s0092-8674(85)80058-1. [DOI] [PubMed] [Google Scholar]
  13. Kamp D., Kahmann R., Zipser D., Broker T. R., Chow L. T. Inversion of the G DNA segment of phage Mu controls phage infectivity. Nature. 1978 Feb 9;271(5645):577–580. doi: 10.1038/271577a0. [DOI] [PubMed] [Google Scholar]
  14. Kitts P. A., Symington L. S., Dyson P., Sherratt D. J. Transposon-encoded site-specific recombination: nature of the Tn3 DNA sequences which constitute the recombination site res. EMBO J. 1983;2(7):1055–1060. doi: 10.1002/j.1460-2075.1983.tb01545.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kreiswirth B. N., Löfdahl S., Betley M. J., O'Reilly M., Schlievert P. M., Bergdoll M. S., Novick R. P. The toxic shock syndrome exotoxin structural gene is not detectably transmitted by a prophage. Nature. 1983 Oct 20;305(5936):709–712. doi: 10.1038/305709a0. [DOI] [PubMed] [Google Scholar]
  16. Krolewski J. J., Murphy E., Novick R. P., Rush M. G. Site-specificity of the chromosomal insertion of Staphylococcus aureus transposon Tn554. J Mol Biol. 1981 Oct 15;152(1):19–33. doi: 10.1016/0022-2836(81)90093-0. [DOI] [PubMed] [Google Scholar]
  17. Landy A., Ross W. Viral integration and excision: structure of the lambda att sites. Science. 1977 Sep 16;197(4309):1147–1160. doi: 10.1126/science.331474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee C. Y., Iandolo J. J. Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene. J Bacteriol. 1986 May;166(2):385–391. doi: 10.1128/jb.166.2.385-391.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lee C. Y., Iandolo J. J. Mechanism of bacteriophage conversion of lipase activity in Staphylococcus aureus. J Bacteriol. 1985 Oct;164(1):288–293. doi: 10.1128/jb.164.1.288-293.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Leong J. M., Nunes-Düby S., Lesser C. F., Youderian P., Susskind M. M., Landy A. The phi 80 and P22 attachment sites. Primary structure and interaction with Escherichia coli integration host factor. J Biol Chem. 1985 Apr 10;260(7):4468–4477. [PubMed] [Google Scholar]
  21. Murphy E., Löfdahl S. Transposition of Tn554 does not generate a target duplication. Nature. 1984 Jan 19;307(5948):292–294. doi: 10.1038/307292a0. [DOI] [PubMed] [Google Scholar]
  22. Murphy E., Phillips S., Edelman I., Novick R. P. Tn554: isolation and characterization of plasmid insertions. Plasmid. 1981 May;5(3):292–305. doi: 10.1016/0147-619x(81)90006-8. [DOI] [PubMed] [Google Scholar]
  23. Novick R. P., Iordanescu S., Surdeanu M., Edelman I. Transduction-related cointegrate formation between Staphylococcal plasmids: a new type of site-specific recombination. Plasmid. 1981 Sep;6(2):159–172. doi: 10.1016/0147-619x(81)90064-0. [DOI] [PubMed] [Google Scholar]
  24. Novick R. P., Projan S. J., Rosenblum W., Edelman I. Staphylococcal plasmid cointegrates are formed by host- and phage-mediated general rec systems that act on short regions of homology. Mol Gen Genet. 1984;195(1-2):374–377. doi: 10.1007/BF00332777. [DOI] [PubMed] [Google Scholar]
  25. Novick R. Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus. Virology. 1967 Sep;33(1):155–166. doi: 10.1016/0042-6822(67)90105-5. [DOI] [PubMed] [Google Scholar]
  26. Pariza M. W., Iandolo J. J. Base ratio and deoxyribonucleic acid homology studies of six Staphylococcus aureus typing bacteriophages. Appl Microbiol. 1974 Feb;27(2):317–323. doi: 10.1128/am.27.2.317-323.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Phillips S., Novick R. P. Tn554--a site-specific repressor-controlled transposon in Staphylococcus aureus. Nature. 1979 Mar 29;278(5703):476–478. doi: 10.1038/278476a0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Plasterk R. H., Van de Putte P. Genetic switches by DNA inversions in prokaryotes. Biochim Biophys Acta. 1984 Jun 16;782(2):111–119. doi: 10.1016/0167-4781(84)90013-7. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Ross W., Landy A. Bacteriophage lambda int protein recognizes two classes of sequence in the phage att site: characterization of arm-type sites. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7724–7728. doi: 10.1073/pnas.79.24.7724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ross W., Landy A., Kikuchi Y., Nash H. Interaction of int protein with specific sites on lambda att DNA. Cell. 1979 Oct;18(2):297–307. doi: 10.1016/0092-8674(79)90049-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ross W., Landy A. Patterns of lambda Int recognition in the regions of strand exchange. Cell. 1983 May;33(1):261–272. doi: 10.1016/0092-8674(83)90355-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  36. Yin S., Bushman W., Landy A. Interaction of the lambda site-specific recombination protein Xis with attachment site DNA. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1040–1044. doi: 10.1073/pnas.82.4.1040. [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