A Global Approach Combining Proteome Analysis and Phenotypic Screening with RNA Interference Yields Novel Apoptosis Regulators

Global approaches like proteome or transcriptome analyses have been performed extensively to identify candidate genes or proteins involved in biological and pathological processes. Here we describe the identification of proteins implicated in the regulation of apoptosis using proteome analysis and t...

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Veröffentlicht in:Molecular & cellular proteomics 2005-01, Vol.4 (1), p.44-55
Hauptverfasser: Machuy, Nikolaus, Thiede, Bernd, Rajalingam, Krishnaraj, Dimmler, Christiane, Thieck, Oliver, Meyer, Thomas F, Rudel, Thomas
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Sprache:eng
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Zusammenfassung:Global approaches like proteome or transcriptome analyses have been performed extensively to identify candidate genes or proteins involved in biological and pathological processes. Here we describe the identification of proteins implicated in the regulation of apoptosis using proteome analysis and the functional validation of targets by RNA interference. A high-throughput platform for the validation of synthetic small interfering RNAs (siRNAs) by quantitative real-time PCR was established. Genes of the identified factors were silenced by automated siRNA transfection, and their role in apoptotic signaling was investigated. Using this strategy, nine new modulators of apoptosis were identified. A subsequent detailed study demonstrated that hepatoma-derived growth factor (HDGF) is required for TNFα-induced release of pro-apoptotic factors from mitochondria. The strategy described here may be used for hypothesis-free, global gene function analysis.
ISSN:1535-9476
1535-9484
1535-9484
DOI:10.1074/mcp.M400089-MCP200