Linked Life Data

12511567

Source:http://linkedlifedata.com/resource/pubmed/id/12511567

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pubmed-article:12511567pubmed:abstractTextSmac/DIABLO is a mitochondrial protein that is proteolytically processed and released during apoptosis along with cytochrome c and other proapoptotic factors. Once in the cytosol, Smac protein binds to inhibitors of apoptosis (IAP) proteins and disrupts the ability of the IAPs to inhibit caspases 3, 7, and 9. The requirement for mitochondrial processing and release has complicated efforts to delineate the effect of Smac overexpression and IAP inhibition on cell death processes. In this report, we document a novel expression system using ubiquitin fusions to express mature, biologically active Smac in the cytosol of transfected cells. Processing of the ubiquitin-Smac fusions is rapid and complete and generates mature Smac protein initiating correctly with the Ala-Val-Pro-Ile tetrapeptide sequence that is required for proper function. The biological activity of this exogenous protein was demonstrated by its interaction with X-linked IAP, one of the most potent of the IAPs. The presence of mature Smac was not sufficient to trigger apoptosis of healthy cells. However, cells with excess Smac protein were greatly sensitized to apoptotic triggers such as etoposide exposure. Cancer cells typically display deregulated apoptotic pathways, including Bcl2 overexpression, thereby suppressing the release of cytochrome c and Smac. The ability to circumvent the requirement for mitochondrial processing and release is critical to developing Smac as a possible gene therapy payload in cancer chemosensitization.lld:pubmed
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pubmed-article:12511567pubmed:articleTitleA novel ubiquitin fusion system bypasses the mitochondria and generates biologically active Smac/DIABLO.lld:pubmed
pubmed-article:12511567pubmed:affiliationSolange Gauthier Karsh Molecular Genetics Laboratory, Children's Hospital of Eastern Ontario, Research Institute, 401 Smyth Road, Ottawa, Ontario K1H 8L1, Canada.lld:pubmed
pubmed-article:12511567pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:12511567pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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