Dynamic interaction between 14-3-3zeta and bax during TNF-a-induced apoptosis in living cells

Xuejuan Gao; Da Xing; Tongsheng Chen

doi:10.1117/12.710927

27 October 2006 Dynamic interaction between 14-3-3zeta and bax during TNF-α-induced apoptosis in living cells

Xuejuan Gao, Da Xing, Tongsheng Chen

Proceedings Volume 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine; 604737 (2006) https://doi.org/10.1117/12.710927
Event: Fourth International Conference on Photonics and Imaging in Biology and Medicine, 2005, Tianjin, China

Abstract

Bax, a proapoptotic member of the Bcl-2 family, localizes largely in the cytoplasm but redistributes to mitochondria and undergoes oligomerization to induce the release of apoptogenic factors such as cytochrome c in response to apoptotic stimuli. Cytoplasmic protein 14-3-3zeta binds to Bax and, upon apoptotic stimulation, releases Bax by a caspase-independent mechanism. However, the direct interaction of the cytoplasmic 14-3-3zeta and Bax in living cells has not been observed. In present study, to monitor the dynamic interaction between 14-3-3zeta and Bax in living cells in real time during apoptosis induced by tumor necrosis factor (TNF-α), DsRed-14-3-3zeta plasmid is constructed. By cotransfecting DsRed- 14-3-3zeta and GFP-Bax plasmids into human lung adenocarcinoma cells (ASTC-a-1), we observe the dynamic interaction between Bax and 14-3-3zeta using fluorescence resonance energy transfer (FRET) technique on laser scanning confocal microscope. The results show that 14-3-3zeta remains in the cytoplasm but GFP-Bax translocates to mitochondria completely after TNF-α stimulation. These results reveal that 14-3-3zeta binds directly to Bax in healthy cells, and that 14-3-3zeta negatively regulates Bax translocation to mitochondria during TNF-α-induced apoptosis.

Citation Download Citation

Xuejuan Gao, Da Xing, and Tongsheng Chen "Dynamic interaction between 14-3-3zeta and bax during TNF-α-induced apoptosis in living cells", Proc. SPIE 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine, 604737 (27 October 2006); https://doi.org/10.1117/12.710927

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