Premium
Conditional Protein Splicing of Inteins from Extremophiles
Author(s) -
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.633.20
Subject(s) - intein , homing endonuclease , protein splicing , thermophile , halophile , rna splicing , hyperthermophile , halobacterium salinarum , genetics , biology , protein engineering , archaea , gene , computational biology , endonuclease , biochemistry , enzyme , bacteria , rna
Protein splicing is a post‐translational process facilitated by an intein, by which the intein catalyzes its own excision from flanking polypeptides (the exteins), concomitant with ligation of the exteins. Many inteins are themselves interrupted by homing endonuclease domains that can facilitate homing into intein‐less alleles. We have discovered inteins from extremophiles that can be expressed in E. coli , purified as unspliced precursors, and induced to splice by cues such as incubation under reducing conditions, at high temperature, or at high ionic strength. We have shown that the activity and folding of inteins from the extreme halophiles Halobacterium salinarum and Haloquadratum walsbyi demonstrate variable dependence on salt concentration, and we have designed transgenic organisms lacking inteins to test the physiological role of splicing. We have solved the structure of an intein from the extreme thermophile Pyrococcus abyssi , and studied the influence of structural elements on its stability at high temperature and pressure. We also have discovered that the splicing and nuclease activity of inteins from extreme thermophiles can be influenced by pressures consistent with those found at the seafloor. Support or Funding Information This work was supported by grants from the National Science Foundation (grants MCB1244089 and MCB1517138) and the Camille and 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 .