A novel locus of <i>Yersinia enterocolitica</i> serotype O:3 involved in lipopolysaccharide outer core biosynthesis

Skurnik, Mikael; Venho, Reija; Toivanen, Paavo; Al-Hendy, Ayman

doi:10.1111/j.1365-2958.1995.mmi_17030575.x

Cited by 126 publications

(170 citation statements)

References 45 publications

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“…The Y. enterocolitica O:3 WT strain 6471/76, its virulence-plasmid-cured derivative 6471/76-c and YeO3-c-R1, a rough derivative of 6471/76-c, were described previously Skurnik, 1984;Skurnik et al, 1995). The strains YeO3-DrfaH, YeO3-c-DrfaH and YeO3-DrfaH/pTM100-rfaH were constructed in this work.…”

Section: Methodsmentioning

confidence: 99%

“…Higher titres of the phages were needed to lyse the YeO3-DrfaH bacteria when compared with the WT bacteria. Phage wR1-37 is specific for the OC hexasaccharide (Pinta et al, 2010;Skurnik et al, 1995), whilst wYeO3-12 is specific for the O-antigen of Y. enterocolitica O:3 (Al-Hendy et al, 1991a; Pajunen et al, 2001).…”

Section: Lps O-antigen and Oc Production Are Rfah-dependentmentioning

confidence: 99%

“…LPS was separated using 4 % stacking and 12 % separating DOC-PAGE as described previously (Krauss et al, 1988;Skurnik et al, 1995;Zhang & Skurnik, 1994). After the run, the material was either visualized by silver staining (Al-Hendy et al, 1991b) or transferred for immunoblotting onto nitrocellulose membrane (Protran, pore size 0.45 mm; Whatman) using the semi-dry apparatus (Owl; Thermo Scientific) as described previously (BiedzkaSarek et al, 2005).…”

Section: Rfah Of Y Enterocolitica O:3mentioning

confidence: 99%

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Expression of the Yersinia enterocolitica O:3 LPS O-antigen and outer core gene clusters is RfaH-dependent

Leskinen

Varjosalo

et al. 2015

Microbiology

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The antiterminator RfaH is required for the expression of LPS, capsule, haemolysin, exotoxin, haemin uptake receptor and F pilus. As these structures are critical for bacterial virulence, loss of RfaH usually leads to attenuation. Here, we inactivated the rfaH gene of Yersinia enterocolitica O:3 to study its role in this enteropathogen. RNA sequencing of the WT and DrfaH strain transcriptomes revealed that RfaH acted as a highly specific regulator that enhanced the transcription of the operons involved in biosynthesis of LPS O-antigen and outer core (OC), but did not affect the expression of enterobacterial common antigen. Interestingly, the transcriptome of the DrfaH strain was very similar to that of an O-antigen-negative mutant. This indicated that some of the changes seen in the DrfaH strain, such as the genes involved in outer membrane homeostasis or in the stress-response-associated Cpx pathway, were actually due to indirect responses via the loss of O-antigen. The decreased amount of LPS on the DrfaH strain cell surface resulted in an attenuated stress response, and lower resistance to compounds such as SDS and polymyxin B. However, the DrfaH strain was significantly more resistant to complement-mediated killing by normal human serum. Taken together, our results revealed a novel role of RfaH acting as a highly specific regulator of O-antigen and OC of LPS in Y. enterocolitica O:3. It may be speculated that RfaH might have an in vivo role in controlling tissue-specific expression of bacterial surface oligo/polysaccharides.

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Section: Methodsmentioning

confidence: 99%

Section: Lps O-antigen and Oc Production Are Rfah-dependentmentioning

confidence: 99%

Section: Rfah Of Y Enterocolitica O:3mentioning

confidence: 99%

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Expression of the Yersinia enterocolitica O:3 LPS O-antigen and outer core gene clusters is RfaH-dependent

Leskinen

Varjosalo

et al. 2015

Microbiology

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“…thermophilus extracellular polysaccharide contains Gal and GalNAc. Y. enterocolitica WbcN was reported to be either a galactosyl-or N-acetylgalactosaminyltransferase (Skurnik et al, 1995). Pseudomonas aeruginosa WbpT was proposed to be an a-N-acetylgalactosaminyltransferase (Belanger et al, 1999) based on sequence similarity and on the presence of the strictly conserved motif EX 7 E, which is present in the carboxy termini of a-D-glycosyltransferases (Geremia et al, 1996).…”

Section: Glycosyltransferasesmentioning

confidence: 99%

The Edwardsiella ictaluri O polysaccharide biosynthesis gene cluster and the role of O polysaccharide in resistance to normal catfish serum and catfish neutrophils

Lawrence

Banes

Azadi³

et al. 2003

Microbiology

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Edwardsiella ictaluri, the causative agent of enteric septicaemia of catfish (ESC), expresses long O polysaccharide (OPS) chains on its surface. The authors previously reported the construction of an isogenic Ed. ictaluri OPS mutant strain and demonstrated that this strain is avirulent in channel catfish. This paper reports the cloning of the Ed. ictaluri OPS biosynthesis gene cluster and identification of the mutated gene in the OPS-negative strain. The sequenced region contains eight complete ORFs and one incomplete ORF encoding LPS biosynthesis enzymes. The mutated gene (designated wbiT ) was similar to other bacterial galactose-4-epimerases. Glycosyl composition analysis indicated that wild-type Ed. ictaluri OPS contains higher amounts of galactose and N-acetylgalactosamine than the OPS mutant strain, which correlated well with predicted functions of the genes identified in the OPS biosynthesis cluster. The OPS mutant had a relatively small, but significant, decrease in its ability to survive in normal catfish serum compared to wild-type Ed. ictaluri, but it retained the ability to resist killing by catfish neutrophils.

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“…These mutants were included in the study as they caused different humoral responses during experimental mouse infections (Skurnik et al, 1995. Based on chemical analyses, YeO3-trs11 and YeO3-trs24 lack OC completely (Rc type), while YeO3-trs22 produces a truncated OC tetrasaccharide (Muszyń ski, 2004).…”

Section: Resultsmentioning

confidence: 99%

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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A novel locus of Yersinia enterocolitica serotype O:3 involved in lipopolysaccharide outer core biosynthesis

Cited by 126 publications

References 45 publications

Expression of the Yersinia enterocolitica O:3 LPS O-antigen and outer core gene clusters is RfaH-dependent

Expression of the Yersinia enterocolitica O:3 LPS O-antigen and outer core gene clusters is RfaH-dependent

The Edwardsiella ictaluri O polysaccharide biosynthesis gene cluster and the role of O polysaccharide in resistance to normal catfish serum and catfish neutrophils

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

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