Date on Honors Thesis
Spring 4-30-2025
Major
Biology
Minor
Chemistry
Examining Committee Member
Dr. Christopher Lennon, Advisor
Examining Committee Member
Dr. Gary ZeRuth, Committee Member
Examining Committee Member
Dr. Chris Trzepacz, Committee Member
Abstract/Description
Inteins are a mobile genetic element hypothesized to function as regulatory molecules in microbes. They bear a homing endonuclease domain for genome traversal, as well as specific residues allowing for protein splicing. In the protein splicing reaction, the intein is removed and the flanking residues, termed exteins, are ligated together with a peptide bond to form a functional host protein. Due to their interruption prior to splicing, inteins prevent a host protein from functioning until the intein ligates the two exteins. To measure factors that regulate protein splicing in vivo, an intein-based biosensor based on kanamycin resistance (Kanamycin Intein Splicing Reporter) was previously developed. In this work, we use our KISR reporter to examine whether chaperones - proteins that assist in the folding of other proteins – can promote the protein splicing reaction. Specifically, we test whether expression of the chaperone GroEL can promote protein splicing within the cellular environment. Indeed, our results demonstrate increased protein splicing increased in the presence of both the KISR and GroEL, but not in the presence of the kanamycin resistance protein lacking the intein and GroEL. Our findings have implications for the potential regulation of protein splicing in nature, as well as on the use of inteins in protein engineering.
Recommended Citation
Thomas, Hazel, "Can Chaperones Promote Protein Splicing?" (2025). Honors College Theses. 283.
https://digitalcommons.murraystate.edu/honorstheses/283