Original Article

Changes in Endoplasmic Reticulum Luminal Environment Affect Cell Sensitivity to Apoptosis

Correspondence to: Marek Michalak, Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7. Tel:(780) 492-2256 Fax:(780) 492-0886 E-mail:marek.michalak{at}ualberta.ca.

To test the role of ER luminal environment in apoptosis, we generated HeLa cell lines inducible with respect to calreticulin and calnexin and investigated their sensitivity to drug-dependent apoptosis. Overexpression of calreticulin, an ER luminal protein, resulted in an increased sensitivity of the cells to both thapsigargin- and staurosporine-induced apoptosis. This correlated with an increased release of cytochrome c from the mitochondria. Overexpression of calnexin, an integral ER membrane protein, had no significant effect on drug-induced apoptosis. In contrast, calreticulin-deficient cells were significantly resistant to apoptosis and this resistance correlated with a decreased release of cytochrome c from mitochondria and low levels of caspase 3 activity. This work indicates that changes in the lumen of the ER amplify the release of cytochrome c from mitochondria, and increase caspase activity, during drug-induced apoptosis. There may be communication between the ER and mitochondria, which may involve Ca²⁺ and play an important role in conferring cell sensitivity to apoptosis. Apoptosis may depend on both the presence of external apoptosis-activating signals, and, as shown in this study, on an internal factor represented by the ER.

Key Words: apoptosis, calreticulin, endoplasmic reticulum, calcium-binding protein

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