Extracellular Vesicles of the Hyperthermophilic Archaeon "Thermococcus onnurineus" NA1T.
Extracellular vesicles (EVs) produced by a sulfur-reducing, hyperthermophilic archaeon, "Thermococcus onnurineus" NA1(T), were purified and characterized. A maximum of four EV bands, showing buoyant densities between 1.1899 and 1.2828 g cm(-3), were observed after CsCl ultracentrifugation. The two major EV bands, B (buoyant density at 25°C [ρ(25)] = 1.2434 g cm(-3)) and C (ρ(25) = 1.2648 g cm(-3)), 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 ∼9.4-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.
Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea.
Dong Hee
DH
Choi
Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea.
Yong Min
YM
Kwon
Marine Microbiology Laboratory, Department of Marine Ecosystems Dynamics, Atmosphere and Ocean Research Institute, University of Tokyo, Kashiwa, Chiba, Japan.
Hiroshi Xavier
HX
Chiura
Marine Ecosystem Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea.
Eun Chan
EC
Yang
Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea Department of Marine Biotechnology, Korea University of Science and Technology, Daejeon, South Korea.
Seung Seob
SS
Bae
Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea Department of Marine Biotechnology, Korea University of Science and Technology, Daejeon, South Korea.
Sung Gyun
SG
Kang
Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea Department of Marine Biotechnology, Korea University of Science and Technology, Daejeon, South Korea.
Jung-Hyun
JH
Lee
Department of Biological Sciences, Sungkyunkwan University, Suwon, South Korea.
Hwan Su
HS
Yoon
Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan, South Korea Department of Marine Biotechnology, Korea University of Science and Technology, Daejeon, South Korea s-jkim@kiost.ac.
Sang-Jin
SJ
Kim
D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D019641:DNA, Archaeal / Q000235:genetics / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D019712:Thermococcus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D019843:Archaeal Proteins; D019641:DNA, Archaeal