Post-translational enzyme activation in an animal via optimized conditional protein splicing.
Control over the timing, location and level of protein activity in vivo is crucial to understanding biological function. Living systems are able to respond to external and internal stimuli rapidly and in a graded fashion by maintaining a pool of proteins whose activities are altered through post-translational modifications. Here we show that the process of protein trans-splicing can be used to modulate enzymatic activity both in cultured cells and in Drosophila melanogaster. We used an optimized conditional protein splicing system to rapidly trigger the in vivo ligation of two inactive fragments of firefly luciferase in a tunable manner. This technique provides a means of controlling enzymatic function with greater speed and precision than with standard genetic techniques and is a useful tool for probing biological processes.
Schwartz
Edmund C
EC
Laboratory of Synthetic Protein Chemistry, The Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Saez
Lino
L
Young
Michael W
MW
Muir
Tom W
TW
D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004330:Drosophila / Q000201:enzymology* / Q000235:genetics; D004789:Enzyme Activation / Q000235:genetics; D047669:Exteins / Q000235:genetics; D047668:Inteins / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D019154:Protein Splicing / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
D017077:Culture Media, Conditioned; D011993:Recombinant Fusion Proteins; D008156:Luciferases