Galactofuranose in Mycoplasma mycoides is important for membrane integrity and conceals adhesins but does not contribute to serum resistance.
2016
Schieck, Elise | Lartigue, Carole | Frey, Joachim | Vozza, Nicolas | Hegermann, Jan | Miller, Rachel A | Valguarnera, Ezequiel | Muriuki, Cecilia | Meens, Jochen | Nene, Vish | Naessens, Jan | Weber, Johann | Lowary, Todd L | Vashee, Sanjay | Feldman, Mario F | Jores, Joerg | International Livestock Research Institute [CGIAR, Nairobi] (ILRI) ; International Livestock Research Institute [CGIAR, Ethiopie] (ILRI) ; Consultative Group on International Agricultural Research [CGIAR] (CGIAR)-Consultative Group on International Agricultural Research [CGIAR] (CGIAR) | Biologie du fruit et pathologie (BFP) ; Université Bordeaux Segalen - Bordeaux 2-Institut National de la Recherche Agronomique (INRA)-Université Sciences et Technologies - Bordeaux 1 (UB) | Universität Bern = University of Bern = Université de Berne (UNIBE) | Department of Biological Sciences ; University of Alberta | Institute of Functional and Applied Anatomy ; Medizinische Hochschule Hannover = Hannover Medical School (MHH) | Department of Food Science ; Cornell University [New York] | Washington University School of Medicine [Saint Louis, MO] | University of Veterinary Medicine [Hannover] | Université de Lausanne = University of Lausanne (UNIL) | University of Alberta | J. Craig Venter Institute [La Jolla, USA] (JCVI)
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Show more [+] Less [-]English. Mycoplasma mycoides subsp. capri (Mmc) and subsp. mycoides (Mmm) are important ruminant pathogens worldwide causing diseases such as pleuropneumonia, mastitis and septicaemia. They express galactofuranose residues on their surface, but their role in pathogenesis has not yet been determined. The M. mycoides genomes contain up to several copies of the glf gene, which encodes an enzyme catalyzing the last step in the synthesis of galactofuranose. We generated a deletion of the glf gene in a strain of Mmc using genome transplantation and tandem repeat endonuclease coupled cleavage (TREC) with yeast as an intermediary host for the genome editing. As expected, the resulting YCp1.1-Δglf strain did not produce the galactofuranose containing glycans as shown by immunoblots and immuno-electronmicroscopy employing a galactofuranose specific monoclonal antibody. The mutant lacking galactofuranose exhibited a decreased growth rate and a significantly enhanced adhesion to small ruminant cells. The mutant was also "leaking" as revealed by a β-galactosidase-based assay employing a membrane impermeable substrate. These findings indicate that galactofuranose-containing polysaccharides conceal adhesins and are important for membrane integrity. Unexpectedly, the mutant strain showed increased serum resistance.
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