Conditional protein splicing of α-sarcin in live cells.
Protein splicing technology harnesses the ability of inteins to ligate protein fragments, forming a mature protein. This report describes our effort to engineer rapamycin-dependent protein splicing of a ribotoxin, called α-sarcin. Engineering this system required the investigation of important splicing parameters, including extein context and splicing temperature. We show α-sarcin splicing is dependent on rapamycin, is inducible with rapid kinetics, and triggers apoptosis in HeLa cells. These findings establish a proof-of-concept for a conditional cell ablation strategy.
Centre for Biomedical Research, University of Victoria, P.O. Box 3020 Station CSC Victoria, BC, Canada V8W 3N5. phoward@uvic.ca.
Spencer C
SC
Alford
Connor
C
O'Sullivan
Jon
J
Obst
Jennifer
J
Christie
Perry L
PL
Howard
D017209:Apoptosis / Q000235:genetics*; D045744:Cell Line, Tumor; D004722:Endoribonucleases / Q000096:biosynthesis / Q000235:genetics*; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D047668:Inteins / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D019154:Protein Splicing / Q000235:genetics*; D020123:Sirolimus / Q000494:pharmacology
D005656:Fungal Proteins; C014801:alpha-sarcin; D049452:Green Fluorescent Proteins; D004722:Endoribonucleases; D020123:Sirolimus