Extracellular Vesicles of the Hyperthermophilic Archaeon “Thermococcus onnurineus” NA1 ^T

Abstract

, were separately purified and counted using a qNano particle analyzer. These EVs, showing different buoyant densities, were identically spherical in shape, and their sizes varied from 80 to 210 nm in diameter, with 120-and 190-nm sizes predominant. The average size of DNA packaged into EVs was about 14 kb. The DNA of the EVs in band C was sequenced and assembled. Mapping of the T. onnurineus NA1 T EV (ToEV) DNA sequences onto the reference genome of the parent archaeon revealed that most genes of T. onnurineus NA1 T were packaged into EVs, except for an ϳ9.4-kb region from TON_0536 to TON_0544. The absence of this specific region of the genome in the EVs was confirmed from band B of the same culture and from bands B and C purified from a different batch culture. The presence of the 3=-terminal sequence and the absence of the 5=-terminal sequence of TON_0536 were repeatedly confirmed. On the basis of these results, we hypothesize that the unpackaged part of the T. onnurineus NA1 T genome might be related to the process that delivers DNA into ToEVs and/or the mechanism generating the ToEVs themselves.

Production of extracellular vesicles with light‐induced proton pump activity by proteorhodopsin‐containing marine bacteria

“…EVs were purified from the 5 L of YIK13 and AKS622 T strain culture medium according to methods previously described (Choi et al, ). Briefly, YIK13 cells grown to the late stationary phase were harvested by centrifugation at 7,500 × g for 40 min at 4°C.…”

“…Remaining cell cultures were centrifuged, filtered through a 0.2 μm syringe filter, and then vesicle samples were resuspended after ultracentrifugation under the same conditions as described above. The fixed cells and EV numbers were analyzed using a qNano (Izon Science, New Zealand) instrument according to a previously described method (Choi et al, ; Kwon et al, ). Cells and EVs were diluted 1,000‐fold into 0.2‐μm filtered 1 × TBT buffer and were measured using NP1000 (500–2000 nm) and NP150 (75–300 nm) membranes, respectively.…”

No genome is an island: toward a 21st century agenda for evolution

“…Other bacterial and archaeal EVs have been found to contain plasmids and viral genomes as well as chromosomal DNA that can function in intercellular DNA transfer . A hyperthermophilic archaeon was particularly interesting because its EVs collectively contained all of the donor genome except for a 9.4 kb region, the absence of which the authors explained as a consequence of the DNA process for delivering DNA to the vesicles …”

“…127,128 A hyperthermophilic archaeon was particularly interesting because its EVs collectively contained all of the donor genome except for a 9.4 kb region, the absence of which the authors explained as a consequence of the DNA process for delivering DNA to the vesicles. 129 Human plasma was found to contain EVs with diverse genomic DNA sequences as cargo. Transfer from plasma EVs into recipient cells was observed in real time after fluorescent staining, and microscopy provided direct evidence that after delivery into the cytosol, EV DNAs could localize inside the nuclear membrane, where they were subsequently documented to undergo transcription and protein expression.…”

Gut microbiota: a new player in regulating immune- and chemo-therapy efficacy

“…EVs are natural nanoparticles delimited by cellular membranous components that carry lipids, carbohydrates, signaling molecules, metabolites, proteins, DNA, RNA, and mediate cell-to-cell communication by delivering their cargo to recipient cells [29] . They are produced and secreted by virtually all cell types in all life kingdoms, including eukaryotic cells [30] , prokaryotic cells [31] , fungi [32] , and archaea [33] . Growing evidence suggests that EVs may represent a universal mechanism of cell-to-cell communication in the same (intrakingdom) or different kingdoms (interkingdom) and regulate gene expression in recipient cells [34] .…”