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Int. J. Mol. Sci. 2018, 19(11), 3510; https://doi.org/10.3390/ijms19113510

A Mutation in the Mesorhizobium loti oatB Gene Alters the Physicochemical Properties of the Bacterial Cell Wall and Reduces Survival inside Acanthamoeba castellanii

1
Department of Genetics and Microbiology, Institute of Microbiology and Biotechnology, Faculty of Biology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland
2
Department of Biochemistry, Institute of Biology, Faculty of Biology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland
3
Department of Comparative Anatomy and Anthropology, Institute of Biology, Faculty of Biology and Biotechnology, Maria Curie-Sklodowska University, Akademicka 19 St., 20-033 Lublin, Poland
*
Author to whom correspondence should be addressed.
Received: 20 September 2018 / Revised: 29 October 2018 / Accepted: 6 November 2018 / Published: 8 November 2018
(This article belongs to the Section Molecular Microbiology)
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Abstract

In our previous report, we had shown that the free-living amoeba Acanthamoeba castellanii influenced the abundance, competiveness, and virulence of Mesorhizobium loti NZP2213, the microsymbiont of agriculturally important plants of the genus Lotus. The molecular basis of this phenomenon; however, had not been explored. In the present study, we demonstrated that oatB, the O-acetyltransferase encoding gene located in the lipopolysaccharide (LPS) synthesis cluster of M. loti, was responsible for maintaining the protective capacity of the bacterial cell envelope, necessary for the bacteria to fight environmental stress and survive inside amoeba cells. Using co-culture assays combined with fluorescence and electron microscopy, we showed that an oatB mutant, unlike the parental strain, was efficiently destroyed after rapid internalization by amoebae. Sensitivity and permeability studies of the oatB mutant, together with topography and nanomechanical investigations with the use of atomic force microscopy (AFM), indicated that the incomplete substitution of lipid A-core moieties with O-polysaccharide (O-PS) residues rendered the mutant more sensitive to hydrophobic compounds. Likewise, the truncated LPS moieties, rather than the lack of O-acetyl groups, made the oatB mutant susceptible to the bactericidal mechanisms (nitrosative stress and the action of lytic enzymes) of A. castellanii. View Full-Text
Keywords: Mesorhizobium loti; Acanthamoeba castellanii; O-acetyltransferase; AFM; cell surface properties; rhizobia-amoebae interactions; lipopolysaccharide Mesorhizobium loti; Acanthamoeba castellanii; O-acetyltransferase; AFM; cell surface properties; rhizobia-amoebae interactions; lipopolysaccharide
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Karaś, M.A.; Turska-Szewczuk, A.; Marczak, M.; Jaszek, M.; Janczarek, M.; Dworaczek, K.; Stefaniuk, D.; Wydrych, J. A Mutation in the Mesorhizobium loti oatB Gene Alters the Physicochemical Properties of the Bacterial Cell Wall and Reduces Survival inside Acanthamoeba castellanii. Int. J. Mol. Sci. 2018, 19, 3510.

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