Method
A pathway sensor for genome-wide screens of intracellular proteolytic cleavage
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* Corresponding author: Brian Seed bseed@ccib.mgh.harvard.edu
1 Center for Computational and Integrative Biology, Massachusetts General Hospital, Cambridge Street, Boston, MA 02114, USA
2 Department of Genetics, Harvard Medical School, Cambridge Street, Boston, MA 02114, USA
3 Origene Technologies Inc., Taft Court, Rockville, MD 20850, USA
Genome Biology 2008, 9:R64 doi:10.1186/gb-2008-9-4-r64
Published: 3 April 2008Abstract
Protein cleavage is a central event in many regulated biological processes. We describe a system for detecting intracellular proteolysis based on non-conventional secretion of Gaussia luciferase (GLUC). GLUC exits the cell without benefit of a secretory leader peptide, but can be anchored in the cell by fusion to β-actin. By including protease cleavage sites between GLUC and β-actin, proteolytic cleavage can be detected. Using this assay, we have identified regulators of autophagy, apoptosis and β-actin cleavage.