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. 1989 Aug;171(8):4146–4153. doi: 10.1128/jb.171.8.4146-4153.1989

Nucleotide sequence and genetic characterization of staphylococcal bacteriophage L54a int and xis genes.

Z H Ye 1, C Y Lee 1
PMCID: PMC210184  PMID: 2526804

Abstract

The nucleotide sequence of a staphylococcal bacteriophage L54a DNA fragment containing genes involved in site-specific recombination was determined. Mutations generated by in vitro mutagenesis were used to map and characterize the int and xis genes. The site-specific recombination functions are tightly clustered within a 1.75-kilobase stretch of DNA fragment with the gene order of attP-int-xis. The int and xis genes are transcribed divergently. The Int protein deduced from the nucleotide sequence has a molecular weight of 41,000. Int is a basic protein with 354 amino acids of which 72 are basic and 38 are acidic. The Xis protein consists of only 59 amino acids with a molecular weight of 7,180. Unlike the Xis proteins of the lambdoid bacteriophages which are all basic proteins, L54a Xis is an acidic protein containing 13 acidic and 8 basic amino acids. The Int protein is required in both integrative and excisive reactions, whereas Xis is only required in excisive reaction. A well-conserved 40-residue region, including three perfectly conserved residues found in 15 site-specific recombinases of the integrase family that have been characterized, was also found in the L54a Int protein.

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Selected References

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

  1. Argos P., Landy A., Abremski K., Egan J. B., Haggard-Ljungquist E., Hoess R. H., Kahn M. L., Kalionis B., Narayana S. V., Pierson L. S., 3rd The integrase family of site-specific recombinases: regional similarities and global diversity. EMBO J. 1986 Feb;5(2):433–440. doi: 10.1002/j.1460-2075.1986.tb04229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bear S. E., Clemens J. B., Enquist L. W., Zagursky R. J. Mutational analysis of the lambda int gene: DNA sequence of dominant mutations. J Bacteriol. 1987 Dec;169(12):5880–5883. doi: 10.1128/jb.169.12.5880-5883.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Davies R. W. DNA sequence of the int-xis-Pi region of the bacteriophage lambda; overlap of the int and xis genes. Nucleic Acids Res. 1980 Apr 25;8(8):1765–1782. doi: 10.1093/nar/8.8.1765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gilbert W. R., Lozeron H. A. Regulation of transcription and DNA replication of bacteriophage phi 80. Virology. 1983 Apr 30;126(2):636–650. doi: 10.1016/s0042-6822(83)80019-1. [DOI] [PubMed] [Google Scholar]
  6. Greenfield L., Simpson L., Kaplan D. Conversion of closed circular DNA molecules to single-nicked molecules by digestion with DNAase I in the presence of ethidium bromide. Biochim Biophys Acta. 1975 Oct 15;407(3):365–375. doi: 10.1016/0005-2787(75)90104-5. [DOI] [PubMed] [Google Scholar]
  7. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [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. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  10. Iordănescu S. Recombinant plasmid obtained from two different, compatible staphylococcal plasmids. J Bacteriol. 1975 Nov;124(2):597–601. doi: 10.1128/jb.124.2.597-601.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Iordănescu S., Surdeanu M. New incompatibility groups of Staphylococcus aureus plasmids. Plasmid. 1980 Nov;4(3):256–260. doi: 10.1016/0147-619x(80)90064-5. [DOI] [PubMed] [Google Scholar]
  12. Iordănescu S., Surdeanu M. Relationships between autonomous and integrated forms of tetracycline resistance plasmid in Staphylococcus aureus. Plasmid. 1979 Apr;2(2):216–224. doi: 10.1016/0147-619x(79)90040-4. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Lacey R. W., Chopra I. Genetic studies of a multi-resistant strain of Staphylococcus aureus. J Med Microbiol. 1974 May;7(2):285–297. doi: 10.1099/00222615-7-2-285. [DOI] [PubMed] [Google Scholar]
  15. Lee C. Y., Buranen S. L. Extent of the DNA sequence required in integration of staphylococcal bacteriophage L54a. J Bacteriol. 1989 Mar;171(3):1652–1657. doi: 10.1128/jb.171.3.1652-1657.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lee C. Y., Iandolo J. J. Integration of staphylococcal phage L54a occurs by site-specific recombination: structural analysis of the attachment sites. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5474–5478. doi: 10.1073/pnas.83.15.5474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Leong J. M., Nunes-Düby S. E., Oser A. B., Lesser C. F., Youderian P., Susskind M. M., Landy A. Structural and regulatory divergence among site-specific recombination genes of lambdoid phage. J Mol Biol. 1986 Jun 20;189(4):603–616. doi: 10.1016/0022-2836(86)90491-2. [DOI] [PubMed] [Google Scholar]
  20. Nakamaye K. L., Eckstein F. Inhibition of restriction endonuclease Nci I cleavage by phosphorothioate groups and its application to oligonucleotide-directed mutagenesis. Nucleic Acids Res. 1986 Dec 22;14(24):9679–9698. doi: 10.1093/nar/14.24.9679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pargellis C. A., Nunes-Düby S. E., de Vargas L. M., Landy A. Suicide recombination substrates yield covalent lambda integrase-DNA complexes and lead to identification of the active site tyrosine. J Biol Chem. 1988 Jun 5;263(16):7678–7685. [PubMed] [Google Scholar]
  22. Rhoads D. D., Roufa D. J. Emetine resistance of Chinese hamster cells: structures of wild-type and mutant ribosomal protein S14 mRNAs. Mol Cell Biol. 1985 Jul;5(7):1655–1659. doi: 10.1128/mcb.5.7.1655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rybchin V. N. Genetics of bacteriophage phi 80--a review. Gene. 1984 Jan;27(1):3–11. doi: 10.1016/0378-1119(84)90233-6. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Susskind M. M., Botstein D. Molecular genetics of bacteriophage P22. Microbiol Rev. 1978 Jun;42(2):385–413. doi: 10.1128/mr.42.2.385-413.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thompson J. F., de Vargas L. M., Skinner S. E., Landy A. Protein-protein interactions in a higher-order structure direct lambda site-specific recombination. J Mol Biol. 1987 Jun 5;195(3):481–493. doi: 10.1016/0022-2836(87)90177-x. [DOI] [PubMed] [Google Scholar]
  29. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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