title: Enterobacterial common antigen and O-specific polysaccharide coexist in the lipopolysaccharide of Yersinia enterocolitica serotype O:3.
abstract: 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 °C and 37 °C 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 ECA-specific 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 °C and also the general reduction in wild-type bacteria of ECA-specific monovalent-reactive material at 37 °C suggested that temperature regulates the expression of ECA. Indeed, RNA-sequencing analysis showed significant downregulation of the ECA biosynthetic gene cluster at 37 °C.
authors.affiliation: Department of Microbiology, University of Silesia, Katowice, Poland.
authors.forename: Artur
authors.initials: A
authors.lastname: Muszyński
authors.affiliation: Department of Microbiology, University of Silesia, Katowice, Poland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Rabsztyn
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute, Research Programs Unit, Immunobiology, University of Helsinki, Helsinki, Finland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Knapska
authors.affiliation: Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), Borstel, Germany.|Department of Microbiology, University of Silesia, Katowice, Poland.
authors.forename: Katarzyna A
authors.initials: KA
authors.lastname: Duda
authors.affiliation: Department of Microbiology, University of Silesia, Katowice, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Duda-Grychtoł
authors.affiliation: Department of Microbiology, University of Silesia, Katowice, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kasperkiewicz
authors.affiliation: Department of Microbiology, University of Silesia, Katowice, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Radziejewska-Lebrecht
authors.affiliation: Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), Borstel, Germany.
authors.forename: Otto
authors.initials: O
authors.lastname: Holst
authors.affiliation: Helsinki University Central Hospital Laboratory Diagnostics, Helsinki, Finland.|Department of Bacteriology and Immunology, Haartman Institute, Research Programs Unit, Immunobiology, University of Helsinki, Helsinki, Finland.
authors.forename: Mikael
authors.initials: M
authors.lastname: Skurnik
mesh_terms: D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D015151:Immunoblotting; D008070:Lipopolysaccharides / Q000737:chemistry* / Q000302:isolation & purification / Q000528:radiation effects; D019081:O Antigens / Q000032:analysis* / Q000302:isolation & purification; D011817:Rabbits; D013696:Temperature; D015008:Yersinia enterocolitica / Q000737:chemistry* / Q000528:radiation effects
chemical_list: D000942:Antigens, Bacterial; D008070:Lipopolysaccharides; D019081:O Antigens; C017286:enterobacterial common antigen