Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki (Jun 2017)
Adaptation of Mycoplasmas to Fluoroquinolones: Modulation of Proteome and Genotoxicity of Extracellular Vesicles of Acholeplasma laidlawii
Abstract
The paper is devoted to the comparative analysis of proteomic profiles and genotoxicity of extracellular vesicles produced by cells that differ in sensitivity to ciprofloxacin of Acholeplasma laidlawii strains – the mycoplasma, being a causative agent of mycoplasmoses of plants and animals, as well as the main contaminant of cell cultures. The relevance of the study is determined by the fact that extracellular vesicles – nanostructures surrounded by a membrane mediating intercellular communication and pathogenesis in bacteria are involved in adaptation of A. laidlawii to antimicrobials and present a new type of infects, the study of which is associated with the prospects of determining the mechanisms of host-parasite systems and solution of problems of pathogen control. The present study has been performed with a view of elucidation of the proteome profile features and assessment of the genotoxicity of extracellular vesicles of A. laidlawii in the development of resistance of the mycoplasma to ciprofloxacin – a drug of fluoroquinolone group which is widely used for inhibiting mycoplasmas. To achieve this goal, we have used the standard microbiological methods, as well as the modern physical and chemical methods, including proteomic profiling with help of 1D-LC-ESI-MS/MS, PCR, automatic scanning, and karyotyping system for assessment of the genotoxicity of vesicles. It has been shown that a significant part of vesicular proteome of A. laidlawii strains with differential sensitivity to ciprofloxacin represents the bacterial virulence factors, as well as that the vesicles of all strains exhibit genotoxicity to lymphocytes of human peripheral blood in vitro. The most important of these results is the fact that the development of resistance of A. laidlawii to ciprofloxacin is accompanied by a significant modulation of vesicular proteome and the increase of mitotoxicity to eukaryotic cells. The obtained data are essential for fundamental research and applied works in the control system of socially significant infections, contaminations of cell cultures and vaccines. The results of the study provide fundamentally new notions about the processes of adaptation of A. laidlawii to antimicrobial drugs and pathogenicity of extracellular vesicles of mycoplasma, as well as suggest the necessity for correction of the control system of mycoplasmas with a view of bacterial vesicles as a new type of infects.
