Cleavage of Armadillo/beta-catenin by the caspase DrICE in Drosophila apoptotic epithelial cells.
BACKGROUND
During apoptosis cells become profoundly restructured through concerted cleavage of cellular proteins by caspases. In epithelial tissues, apoptotic cells loose their apical/basal polarity and are extruded from the epithelium. We used the Drosophila embryo as a system to investigate the regulation of components of the zonula adherens during apoptosis. Since Armadillo/beta-catenin (Arm) is a major regulator of cadherin-mediated adhesion, we analyzed the mechanisms of Arm proteolysis in apoptosis.


RESULTS
We define early and late apoptotic stages and find that early in apoptosis Dalpha-catenin remains relatively stable, while Arm and DE-cadherin protein levels are strongly reduced. Arm is cleaved by caspases in embryo extracts and we provide evidence that the caspase-3 homolog drICE cleaves Arm in vitro and in vivo. Cleavage by drICE creates a stable protein fragment that remains associated with the plasma membrane early in apoptosis. To further understand the role of caspase-mediated cleavage of Arm, we examined potential caspase cleavage sites and found that drICE cleaves Arm at a unique DQVD motif in the N-terminal domain of the protein. Mutation of the drICE cleavage site in Arm results in a protein that is not cleaved in vitro and in vivo. Furthermore we provide evidence that cleavage of Arm plays a role in the removal of DE-cadherin from the plasma membrane during apoptosis.


CONCLUSIONS
This study defines the specificity of caspase cleavage of Arm in Drosophila apoptotic cells. Our data suggest that N-terminal truncation of Arm by caspases is evolutionarily conserved and thus might provide a principal mechanism involved in the disassembly of adherens junctions during apoptosis.
Kessler
Thomas
T
Institut für Genetik, Heinrich Heine Universität, Düsseldorf, Germany. kessler@molgen.mpg.de
Müller
H Arno J
HA
D022005:Adherens Junctions / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D051186:Armadillo Domain Proteins / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism; D020169:Caspases / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004330:Drosophila / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D051178:alpha Catenin / Q000378:metabolism
C495305:ARM protein, Drosophila; D051186:Armadillo Domain Proteins; D015820:Cadherins; D015815:Cell Adhesion Molecules; D029721:Drosophila Proteins; D014157:Transcription Factors; D051178:alpha Catenin; C081439:alpha-Cat protein, Drosophila; C501889:shg protein, Drosophila; D020169:Caspases; C109171:drICE protein, Drosophila