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Tissue distribution of Diablo/Smac revealed by monoclonal antibodies
1The Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, VIC 3050, Australia. tikoo@wehi.edu.au
Abstract:
Diablo/Smac is a mammalian pro-apoptotic protein that can antagonize the inhibitor of apoptosis proteins (IAPs). We have produced monoclonal antibodies specific for Diablo and have used these to examine its tissue distribution and subcellular localization in healthy and apoptotic cells. Diablo could be detected in a wide range of mouse tissues including liver, kidney, lung, intestine, pancreas and testes by Western blot analysis. Immunohistochemical analysis found Diablo to be most abundant in the germinal cells of the testes, the parenchymal cells of the liver and the tubule cells of the kidney. In support of previous subcellular localization analysis, Diablo was present within the mitochondria of healthy cells, but released into the cytosol following the induction of apoptosis by UV.
Insights
Diablo/Smac, a pro-apoptotic protein, is found in various mouse tissues. Its location shifts from mitochondria to cytosol during apoptosis, indicating its role in programmed cell death.
Area of Science:
- Cell Biology
- Molecular Biology
- Immunology
Background:
- Diablo/Smac is a mammalian pro-apoptotic factor.
- It antagonizes inhibitor of apoptosis proteins (IAPs).
Purpose of the Study:
- To produce monoclonal antibodies specific for Diablo/Smac.
- To examine Diablo/Smac tissue distribution and subcellular localization in healthy and apoptotic cells.
Main Methods:
- Western blot analysis for tissue distribution.
- Immunohistochemical analysis for subcellular localization.
- UV-induced apoptosis model.
Main Results:
- Diablo/Smac detected in liver, kidney, lung, intestine, pancreas, and testes.
- Highest abundance in testes germinal cells, liver parenchymal cells, and kidney tubule cells.
- Diablo/Smac localized to mitochondria in healthy cells, released to cytosol upon UV-induced apoptosis.
Conclusions:
- Monoclonal antibodies specific for Diablo/Smac were successfully produced.
- Diablo/Smac exhibits wide tissue distribution and specific subcellular localization.
- Diablo/Smac translocation from mitochondria to cytosol is a key event in UV-induced apoptosis.