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Under Pressure: Pressure and Temperature Dependent Nuclease Activity
Author(s) -
Wibben Amanda C,
Comeau Hannah Y,
Mills Kenneth V
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.633.31
Subject(s) - homing endonuclease , intein , endonuclease , thermophile , archaea , nuclease , hydrostatic pressure , extremophile , hydrothermal vent , biology , dna , enzyme , genetics , chemistry , biochemistry , gene , hydrothermal circulation , physics , rna , rna splicing , thermodynamics , paleontology
An intein is an intervening protein segment that can be found in proteins from extremophiles and is typically self‐excised to allow for complete flanking protein activity. We are interested in studying specific inteins that contain a homing endonuclease (HE), an enzyme that cleaves double stranded DNA in order to initiate insertion of an intein gene by homologous recombination. We are focused on the homing endonuclease activity of inteins from two hyperthermophilic archaea: Thermococcus barophilus (Tba) , isolated from a deep sea hydrothermal vent, and Thermococcus kodakarensis (Tko) , isolated from a volcanic area on the shore of a Japanese island. Because these intein‐containing extremophiles have been isolated from very high temperatures but from different depths, we are interested in the differential homing endonuclease activity of these extremophiles at varied temperatures and pressures. We have shown that Tba HE is active at pressures up to 1000 atm. Given that Tko is a surfaced‐welling organism, we suspect that the Tko HE will be pressure sensitive while Tba, which lives naturally at a pressure near 400 atm, is not. Additionally, we have found that both Tba HE and Tko HE show temperature dependence, which we attribute to the extreme temperatures that both organisms reside in. Support or Funding Information This work was supported by the National Science Foundation (grant MCB‐1517138) and by the Camille & Henry Dreyfus Foundation. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .