Disruption of occludin function in polarized epithelial cells activates the extrinsic pathway of apoptosis leading to cell extrusion without loss of transepithelial resistance.
BACKGROUND
Occludin is a tetraspanin protein normally localized to tight junctions. The protein interacts with a variety of pathogens including viruses and bacteria, an interaction that sometimes leads to its extrajunctional localization.


RESULTS
Here we report that treatment of mammary epithelial monolayers with a circularized peptide containing a four amino acid sequence found in the second extracellular loop of occludin, LHYH, leads to the appearance of extrajunctional occludin and activation of the extrinsic apoptotic pathway. At early times after peptide treatment endogenous occludin and the LYHY peptide were co-localized in extrajunctional patches, which were also shown to contain components of the death inducing signaling complex (DISC), caspases 8 and 3, the death receptor FAS and the adaptor molecule FADD. After this treatment occludin could be immunoprecipitated with FADD, confirming its interaction with the DISC. Extrusion after LYHY treatment was accomplished with no loss of epithelial resistance.


CONCLUSIONS
These observations provide strong evidence that, following disruption, occludin forms a complex with the extrinsic death receptor leading to extrusion of apoptotic cells from the epithelial monolayer. They suggest that occludin has a protective as well as a barrier forming role in epithelia; pathogenic agents which utilize this protein as an entry point into the cell might set off an apoptotic reaction allowing extrusion of the infected cell before the pathogen can gain entry to the interstitial space.
Beeman
Neal E
NE
Department of Physiology and Biophysics, School of Medicine, University of Colorado Denver, Aurora, CO 80045, USA.
Baumgartner
Heidi K
HK
Webb
Patricia G
PG
Schaack
Jerome B
JB
Neville
Margaret C
MC
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement; D016764:Cell Polarity; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D062793:Occludin; D011485:Protein Binding; D053218:Receptors, Death Domain / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism
D008565:Membrane Proteins; D062793:Occludin; C568986:Ocln protein, mouse; D053218:Receptors, Death Domain; D020169:Caspases