A pathway sensor for genome-wide screens of intracellular proteolytic cleavage.
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 beta-actin. By including protease cleavage sites between GLUC and beta-actin, proteolytic cleavage can be detected. Using this assay, we have identified regulators of autophagy, apoptosis and beta-actin cleavage.
Ketteler
Robin
R
Center for Computational and Integrative Biology, Massachusetts General Hospital, Cambridge Street, Boston, MA 02114, USA.
Sun
Zairen
Z
Kovacs
Karl F
KF
He
Wei-Wu
WW
Seed
Brian
B
D000199:Actins / Q000378:metabolism*; D017209:Apoptosis; D001343:Autophagy; D010450:Endopeptidases / Q000032:analysis / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D011993:Recombinant Fusion Proteins
D000199:Actins; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D010450:Endopeptidases