ECA-Antibodies in Antisera Against R Mutants of Yersinia enterocolitica O:3

Radziejewska‐Lebrecht, Joanna; Kasperkiewicz, Katarzyna; Skurnik, Mikael; Brade, Lore; Steinmetz, Ivo; Świerzko, Anna St.; Muszyński, Artur

doi:10.1007/0-306-48416-1_42

Cited by 16 publications

(25 citation statements)

References 3 publications

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“…On the other hand, incubation of bacteria with periodate clearly destroyed the phage receptor. These results thus strengthened the conclusion that the outer core hexasaccharide structure (Radziejewska-Lebrecht et al, 1998) is the phage receptor.…”

Section: Characterization Of Wr1-37 Receptorsupporting

confidence: 67%

Yersiniophage ϕR1-37 is a tailed bacteriophage having a 270 kb DNA genome with thymidine replaced by deoxyuridine

et al. 2005

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Yersiniophage wR1-37 is a tailed bacteriophage having a 270 kb DNA genome with thymidine replaced by deoxyuridine Bacteriophage wR1-37 was isolated based on its ability to infect strain YeO3-R1, a virulenceplasmid-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 wR1-37 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. wR1-37 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 wR1-37 genome was dsDNA and~270 kb in size. Enzymically hydrolysed DNA was subjected to HPLC-MS/MS analysis, which demonstrated that the wR1-37 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 wR1- 37 have not yet been characterized. Genes for two of the structural proteins, p24 and p46, were identified from the partially sequenced wR1-37 genome. INTRODUCTIONYersinia enterocolitica is a Gram-negative bacterium of the family Enterobacteriaceae. The species includes over 30 serotypes, a number of which are human pathogens (Wauters et al., 1991). While the major pathogenic serotypes in Europe, Canada, Japan and South Africa are O : 3 and O : 9; in the United States serotype O : 8 is more prevalent. Y. enterocolitica is widely distributed in nature, swine being the major reservoir of the pathogenic strains (Bottone, 1997).Bacteriophages are the most abundant organisms on Earth, and it is estimated that for each microbial isolate at least 10 different phages can be found (Hendrix, 2002;Pedulla et al., 2003). A number of phages infecting the members of the genus Yersinia have been isolated (Baker & Farmer, 1982;Popp et al., 2000;Stevenson & Airdrie, 1984), but relatively few have been characterized in more detail. Y. enterocolitica O : 3-specific phage wYeO3-12 has been shown to be closely related to Escherichia coli phages T3 and T7 (Pajunen et al., 2000(Pajunen et al., , 2001. Likewise, Yersinia pestis phage wA1122 was recently found to be a close relative of T7 (Garcia et al., 2003a). Bacteriophage...

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Section: Characterization Of Wr1-37 Receptorsupporting

confidence: 67%

Yersiniophage ϕR1-37 is a tailed bacteriophage having a 270 kb DNA genome with thymidine replaced by deoxyuridine

et al. 2005

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“…Our previous work clearly demonstrated the presence of ECA PG and ECA LPS in the S-type LPS of Ye75S and in Rmutants (Rabsztyn et al, 2011;Radziejewska-Lebrecht et al, 1998, 2003 and that expression of ECA PG and ECA LPS is decreased at 37 u C (Muszyń ski, 2004;Rabsztyn et al, 2011). In the current study we clearly show a downregulation of the expression of ECA LPS at 37 u C compared to ECA LPS of bacteria grown at 22 uC.…”

Section: Eca and Ops Coexist In The Lps Of Y Enterocolitica O :supporting

confidence: 75%

“…1) Skurnik et al, 1999). We previously demonstrated the presence of ECA LPS in Y. enterocolitica O : 3 wild-type strains and in R-mutants expressing full or truncated OC (Rabsztyn et al, 2011;Radziejewska-Lebrecht et al, 1998, 2003.…”

Section: Introductionmentioning

confidence: 94%

“…The third form of ECA, found in the periplasmic space, is a cyclic form and has no lipid anchor (ECA CYC ) (Kajimura et al, 2005;Kuhn et al, 1988;Lugowski et al, 1983). Of these three ECA types, only ECA LPS is naturally immunogenic and it has been detected from Escherichia coli, Proteus mirabilis and Yersinia enterocolitica Kasperkiewicz et al, 2004;Kuhn et al, 1988;Rabsztyn et al, 2011;Radziejewska-Lebrecht et al, 2003). Although the role of ECA in Enterobacteriaceae is not fully understood, there is growing evidence for its protective function against environmental stress.…”

Section: Introductionmentioning

confidence: 99%

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Enterobacterial common antigen and O-specific polysaccharide coexist in the lipopolysaccharide of Yersinia enterocolitica serotype O : 3

et al. 2013

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Yersinia enterocolitica serotype O : 3 produces two types of lipopolysaccharide (LPS) molecules to its surface. In both types the lipid A (LA) structure is substituted by inner core (IC) octasaccharide to which either outer core (OC) hexasaccharide or homopolymeric O-polysaccharide (OPS) is linked. In addition, enterobacterial common antigen (ECA) can be covalently linked to LPS, however, via an unknown linkage. To elucidate the relationship between ECA and LPS in Y. enterocolitica O : 3 and the effect of temperature on their expression, LPS was isolated from bacteria grown at 22 6C and 37 6C by consequent hot phenol/water and phenol-chloroform-light petroleum extractions to obtain LPS preparations free of ECA linked to glycerophospholipid. In immunoblotting, monoclonal antibodies TomA6 and 898, specific for OPS and ECA, respectively, reacted both with ladder-like bands and with a slower-migrating smear suggesting that the ECA and OPS epitopes coexist on the same molecules. These results were supported by immunoblotting with a monovalent Y. enterocolitica O : 3 ECAspecific rabbit antiserum. Also, two or three 898-positive (and monovalent-positive) TomA6-negative bands migrated at the level of the LA-IC band in LPS samples from certain OC mutants, most likely representing LA-IC molecules carrying 1-3 ECA repeat units but no OPS. These bands were also present in Y. enterocolitica O : 9 OC mutants; however, coexistence of ECA and OPS in the same molecules could not be detected. Finally, the LA-IC-ECA bands were missing from LPS of bacteria grown at 37 6C and also the general reduction in wild-type bacteria of ECA-specific monovalentreactive material at 37 6C suggested that temperature regulates the expression of ECA. Indeed, RNAsequencing analysis showed significant downregulation of the ECA biosynthetic gene cluster at 37 6C.

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“…In subsequent studies, Kunin et al (9) and Mayer et al (20) identified the presence of such an immunogenic form of ECA in rough mutants of E. coli Ra, R1, R4, and K-12 that express a complete core OS region of LOS. More recent and extensive serological studies have addressed the occurrence of ECA LPS in Yersinia enterocolitica (21)(22)(23)(24) and Proteus mirabilis. ECA immunogenicity has also been demonstrated for several wild-type strains and mutants of Y. enterocolitica O:3 and O:9 as well as P. mirabilis (25).…”

Section: Enterobacterial Common Antigen (Eca)mentioning

confidence: 99%

First Evidence for a Covalent Linkage between Enterobacterial Common Antigen and Lipopolysaccharide in Shigella sonnei Phase II ECALPS

Gozdziewicz

Ługowski

Łukasiewicz

2014

Journal of Biological Chemistry

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Background: Enterobacterial common antigen (ECA) is a surface antigen of all enteric bacteria. Results: ECA polysaccharide substitutes the outer core region of Shigella sonnei lipopolysaccharide (LPS). Conclusion: First structural evidence for the existence of ECA covalently associated with LPS (ECA LPS ). Significance: ECA LPS is the only immunogenic form of ECA and could be a target for therapeutic strategies against nosocomial infections.

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ECA-Antibodies in Antisera Against R Mutants of Yersinia enterocolitica O:3

Cited by 16 publications

References 3 publications

Yersiniophage ϕR1-37 is a tailed bacteriophage having a 270 kb DNA genome with thymidine replaced by deoxyuridine

Yersiniophage ϕR1-37 is a tailed bacteriophage having a 270 kb DNA genome with thymidine replaced by deoxyuridine

Enterobacterial common antigen and O-specific polysaccharide coexist in the lipopolysaccharide of Yersinia enterocolitica serotype O : 3

First Evidence for a Covalent Linkage between Enterobacterial Common Antigen and Lipopolysaccharide in Shigella sonnei Phase II ECALPS

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