Galactofuranose in Mycoplasma mycoides is important for membrane integrity and conceals adhesins but does not contribute to serum resistance.
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 catalysing 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.
International Livestock Research Institute, Old Naivasha Road, P.O. Box 30709, 00100, Nairobi, Kenya.
Elise
E
Schieck
UMR 1332 Biologie du Fruit et Pathologie, The French National Institute for Agricultural Research, INRA-Université Bordeaux, Segalen, 71, avenue Edouard Bourlaux, CS20032, F-33882, Villenave D'Ornon CEDEX, Bordeaux, France.|UMR 1332 de Biologie du Fruit et Pathologie, Université Bordeaux, F-33140, Villenave d'Ornon, Bordeaux, France.
Carole
C
Lartigue
Institute of Veterinary Bacteriology, University of Bern, CH-3001, Bern, Switzerland.
Joachim
J
Frey
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada.
Nicolas
N
Vozza
Institute of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany.|Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), Hannover, Germany.|REBIRTH Cluster of Excellence, Hannover, Germany.
Jan
J
Hegermann
Department of Food Science, Cornell University, Ithaca, NY, USA.
Rachel A
RA
Miller
Department of Molecular Microbiology, Washington University School of Medicine St Louis, 660 South Euclid Avenue, St Louis, MO 63110, USA.
Ezequiel
E
Valguarnera
International Livestock Research Institute, Old Naivasha Road, P.O. Box 30709, 00100, Nairobi, Kenya.
Cecilia
C
Muriuki
Institute for Microbiology, Department of Infectious Diseases, University of Veterinary Medicine Hannover, Hannover, Germany.
Jochen
J
Meens
International Livestock Research Institute, Old Naivasha Road, P.O. Box 30709, 00100, Nairobi, Kenya.
Vish
V
Nene
International Livestock Research Institute, Old Naivasha Road, P.O. Box 30709, 00100, Nairobi, Kenya.
Jan
J
Naessens
Center for Integrative Genomics, Lausanne Genomic Technologies Facility,University of Lausanne, Lausanne, Switzerland.
Johann
J
Weber
Department of Chemistry, Alberta Glycomics Centre, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada.
Todd L
TL
Lowary
J. Craig Venter Institute, 9704 Medical Center Drive, MD 20850, Rockville, USA.
Sanjay
S
Vashee
Department of Biological Sciences, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada.|Department of Molecular Microbiology, Washington University School of Medicine St Louis, 660 South Euclid Avenue, St Louis, MO 63110, USA.
Mario F
MF
Feldman
International Livestock Research Institute, Old Naivasha Road, P.O. Box 30709, 00100, Nairobi, Kenya.|Institute of Veterinary Bacteriology, University of Bern, CH-3001, Bern, Switzerland.
Joerg
J
Jores
D018829:Adhesins, Bacterial / Q000032:analysis*; D000818:Animals; D001422:Bacterial Adhesion; D001770:Blood Bactericidal Activity; D002462:Cell Membrane / Q000737:chemistry* / Q000502:physiology*; D002478:Cells, Cultured; D004187:Disaccharides / Q000032:analysis*; D017353:Gene Deletion; D018390:Gene Targeting; D015151:Immunoblotting; D016253:Microscopy, Immunoelectron; D009176:Mycoplasma mycoides / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D012756:Sheep
D018829:Adhesins, Bacterial; D004187:Disaccharides