A pathway sensor for genome-wide screens of intracellular proteolytic cleavage

Robin Ketteler¹^,2 , Zairen Sun³ , Karl F Kovacs³ , Wei-Wu He³ and Brian Seed¹^,2

¹Center for Computational and Integrative Biology, Massachusetts General Hospital, Cambridge Street, Boston, MA 02114, USA

²Department of Genetics, Harvard Medical School, Cambridge Street, Boston, MA 02114, USA

³Origene Technologies Inc., Taft Court, Rockville, MD 20850, USA

author email corresponding author email

Genome Biology 2008, 9:R64doi:10.1186/gb-2008-9-4-r64


Published:	3 April 2008

Subject areas: Cell biology, Methods

Abstract

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.