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Microbiology 151 (2005), 4093-4102; DOI  10.1099/mic.0.28265-0
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Microbiology 151 (2005), 4093-4102; DOI  10.1099/mic.0.28265-0
© 2005 Society for General Microbiology

Yersiniophage {phi}R1-37 is a tailed bacteriophage having a 270 kb DNA genome with thymidine replaced by deoxyuridine

Saija Kiljunen1,4, Kristo Hakala2,3, Elise Pinta1,4, Suvi Huttunen1,{dagger}, Patrycja Pluta1, Aneta Gador1, Harri Lönnberg2 and Mikael Skurnik1,4

1 Department of Medical Biochemistry and Molecular Biology, Institute of Biomedicine, University of Turku, Turku, Finland
2 Department of Chemistry, University of Turku, Turku, Finland
3 Department of Pharmacology and Clinical Pharmacology, Institute of Biomedicine, University of Turku, Turku, Finland
4 Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki and Helsinki University Central Hospital Laboratory, Helsinki, Finland

Correspondence
Saija Kiljunen
saija.kiljunen{at}utu.fi

Bacteriophage {phi}R1-37 was isolated based on its ability to infect strain YeO3-R1, a virulence-plasmid-cured O antigen-negative derivative of Yersinia enterocolitica serotype O : 3. In this study, the phage receptor was found to be a structure in the outer core hexasaccharide of Y. enterocolitica O : 3 LPS. The phage receptor was present in the outer core of strains of many other Y. enterocolitica serotypes, but also in some Yersinia intermedia strains. Surprisingly, the receptor structure resided in the O antigen of Yersinia pseudotuberculosis O : 9. Electron microscopy demonstrated that {phi}R1-37 particles have an icosahedral head of 88 nm, a short neck of 10 nm, a long contractile tail of 236 nm, and tail fibres of at least 86 nm. This implies that the phage belongs to the order Caudovirales and the family Myoviridae in the ICTV (International Committee for Taxonomy of Viruses) classification. {phi}R1-37 was found to have a lytic life cycle, with eclipse and latent periods of 40 and 50 min, respectively, and a burst size of ~80 p.f.u. per infected cell. Restriction digestions and PFGE showed that the {phi}R1-37 genome was dsDNA and ~270 kb in size. Enzymically hydrolysed DNA was subjected to HPLC-MS/MS analysis, which demonstrated that the {phi}R1-37 genome is composed of DNA in which thymidine (T) is >99 % replaced by deoxyuridine (dU). The only organisms known to have similar DNA are the Bacillus subtilis-specific bacteriophages PBS1 and PBS2. N-terminal amino acid sequences of four major structural proteins did not show any similarity to (viral) protein sequences in databases, indicating that close relatives of {phi}R1-37 have not yet been characterized. Genes for two of the structural proteins, p24 and p46, were identified from the partially sequenced {phi}R1-37 genome.


Abbreviations: BER, base-excision repair pathway; DOC, deoxycholate; ICTV, International Committee for Taxonomy of Viruses; RT, room temperature; Ugi, uracil-DNA glycosylase inhibitor; Ung, uracil-DNA glycosylase

The GenBank/EMBL/DDBJ accession numbers for the sequences reported in this paper are AJ972879 and AJ972880.

{dagger}Present address: Department of Virology, University of Turku, Tykistokatu 6 A, 20520 Turku, Finland.




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