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Etude des vésicules membranaires produites par les Archées hyperthermophiles marines de l’ordre des Thermococcales

Abstract : Secretion of membrane vesicles (MVs) is an important physiological process that has been extensively studied in Bacteria and Eukarya. The recent discovery that Archaea produce MVs shows that this process is universal and suggests that the Last Universal Common Ancestor, LUCA, certainly produced MVs. As these archaeal MVs have been only studied in some Crenarchaeota (ex: G/ Sulfolobus), we started characterizing MVs produced by Thermococcales, a group of hyperthermophilic anaerobic Euryarchaeota.In the first part of this study we examined the mechanism of production as well as the protein and lipid composition of MVs produced by three strains of Thermococcales: Thermococcus kodakaraensis, Thermococcus gammatolerans and Thermocococus sp. 5-4. We observed that MVs are released by a budding process from the cell envelope that is similar to ectosome formation in eukaryotic cells. Moreover, clusters of MVs often form filamentous structures and protuberances on cell surfaces, resembling recently described bacterial nanopods. Differences in structure are observable between MVs of the three species, as well as in their protein composition. However, MVs and cell membranes from the same species have a quite similar protein and lipid composition, confirming that MVs are produced from cell membranes. A major protein present in cell membranes and MVs from the three strains is the oligopeptide-binding proteins (OppA), which has homologues in MVs from Sulfolobus species. Thermococcales MVs harbor DNA and protect this DNA against thermodegradation. Here, we show that T. kodakaraensis cells transformed with the shuttle plasmid pLC70 release MVs harboring this plasmid. Interestingly, these MVs can be used to transfer pLC70 into plasmid-free cells, suggesting that MVs could be involved in DNA transfer between cells at high temperature. In the second part of this study, we were specially interested in the strain Thermococcus nautilus, a Thermococcale that produces MVs selectively enriched in two plasmids from the cell. Notably, one of them corresponds to the genome of a defective virus from PRD1-adenovirus lineage. This indicates that MVs can be used as vehicles for the transport of viral genomes and suggests that production of MVs by ancestral cells could have played a role in the origin of viruses.In addition to be involved in transport of plasmids/viruses, MVs from T. nautilus display a toxic effect on some strains of Thermococcales, maybe due to the delivery of toxins. Even if these “thermococcins” remain to be characterized, this is the first time that a toxic activity associated with MVs has been shown in Thermococcales.
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Marie Gaudin. Etude des vésicules membranaires produites par les Archées hyperthermophiles marines de l’ordre des Thermococcales. Sciences agricoles. Université Paris Sud - Paris XI, 2012. Français. ⟨NNT : 2012PA112093⟩. ⟨tel-00716699⟩

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