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Visit our Expo - Redox and Inflammation signaling 2012

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Session X : Cell death in cancer Poster X, 20<br />

Relationship between apoptotic <strong>and</strong> autophagic programmed cell death: role of Bcl-2<br />

<strong>and</strong> Beclin 1 proteins<br />

Iwona A. Ciechomska, Christoph Goemans, Aviva Tolkovsky<br />

Department of Biochemistry, University of Cambridge, Tennis C<strong>our</strong>t Road, Cambridge,<br />

CB2 1QW, UK<br />

Email: iac23@mole.bio.cam.ac.uk; cgg20@mole.bio.cam.ac.uk;<br />

amt@mole.bio.cam.ac.uk<br />

Programmed cell death (PCD) mechanisms can be divided into several categories including<br />

type I (apoptosis) <strong>and</strong> type II (autophagic cell death). The mutual relationship between<br />

apoptotic <strong>and</strong> autophagic death is currently debated because Beclin 1, an important regulator<br />

of autophagy, is also a Bcl-2 interacting protein. Recent studies revealed that the interaction<br />

between Bcl-2 <strong>and</strong> Beclin 1 regulates starvation-induced autophagy.<br />

In this study we investigated how the interactions between Bcl-2 <strong>and</strong> Beclin-1 influence the<br />

apoptotic machinery. In order to investigate this effect first we have studied how each of these<br />

proteins individually, Bcl-2 or Beclin 1, influence the level of apoptosis in HeLa cells induced<br />

by a variety of apoptotic stimuli. The Bcl-2 protein is localized in the different cellular<br />

membranes, therefore wild-type <strong>and</strong> Bcl-2 variants that are exclusively targeted to the<br />

endoplasmic reticulum (ER) membrane or the mitochondrial outer membrane (MOM) were<br />

used. We have confirmed that Bcl-2 decreases the level of apoptosis induced by staurosporine<br />

independently on the localization, although we have found some toxic effect of wild-type Bcl-<br />

2 after transient overexpression in untreated cells. Overexpression of either wild-type or<br />

Beclin 1 mutants that cannot bind to Bcl-2 resulted in increased apoptosis. Furthermore, Bcl-2<br />

protein binds to Beclin 1 <strong>and</strong> the intracellular localization of Beclin 1 was changed according<br />

to Bcl-2 localisation. Next we examined how the interaction between Beclin 1 <strong>and</strong> Bcl-2<br />

influence the level of apoptosis. Beclin 1 mutants that are defective in Bcl-2 binding induced<br />

less apoptosis than wild-type Beclin 1 after overexpression of ER- or MOM-targeted Bcl-2.<br />

Thus, <strong>our</strong> data suggest that Beclin 1 not only functions as a proautophagy protein, but also as<br />

a proapoptotic protein via disruption of the anti-apoptotic function of Bcl-2.<br />

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