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Differential Protein Splicing of Salt Dependent Inteins from Haloquadratum Walsybi
Author(s) -
Lynch Abigeal Kathleen,
Amunya Shirley,
Reitter Julie,
Mills Kenneth
Publication year - 2017
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.31.1_supplement.606.13
Subject(s) - intein , protein splicing , rna splicing , splice , fusion protein , chemistry , trans splicing , in vitro , biochemistry , biology , recombinant dna , gene , rna
Protein splicing is a self‐catalyzed, post‐translational process in which an intervening polypeptide (intein) catalyzes its own removal from the flanking polypeptides (exteins), as well as the ligation of the exteins. We are interested in the splicing of two inteins interrupting the cdc21 protein from Haloquadratum walsbyi ( Hwa ). Hwa is a halophilic archaeon. Because of this, we hypothesized that protein splicing would be salt dependent. The first intein, Cdc21a, does not splice on overexpression in E. coli, but can be induced to splice in vitro on incubation with 2.5 M sodium chloride at 28°C. The second intein, Cdc21d, splices in vivo , and further splicing can be induced in vitro with higher concentrations of salt. Although the first intein requires 2.5 M salt to splice, Cdc21d requires only 500 mM salt to induce additional splicing. Given that both inteins interrupt the same protein, this differential activity could result in different protein products due to alternative splicing in the full precursor. We are studying this relative activity when the two inteins are expressed in the same precursor fusion protein. Support or Funding Information This work was supported by the National Science Foundation (grants MCB‐1244089 and MCB‐1517138 to KVM) and the Dreyfus Foundation (KVM).