New Approach to Identify Novel Regulators of Myc Oncoprotein Stability.

摘要

The Myc oncoprotein is deregulated in the majority of breast cancers yet it has not been possible to develop a therapeutic to target Myc using traditional approaches. It has recently been shown that targeting Myc for degradation may offer a new therapeutic strategy to decrease Myc levels and kill tumor cells that are addicted to this potent oncogene. Identifying Myc protein regulatory mechanisms has been a time and labor-intensive process using classical methods to measure endogenous Myc half-life. Indeed, only one pathway has been identified to date. To facilitate this research our concept was that we could build a Myc stability probe by generating an in-frame fusion protein of Myc with a fluorescent protein (Myc-FP). After de novo synthesis, some FPs require hours to fold properly before they will exhibit fluorescence. Our concept was that if Myc-FP was turned over every 30 min, like Myc, then the short half-life Myc-FP would not demonstrate fluorescence, but if Myc was deregulated at the level of protein turnover, then Myc-FP would exhibit fluorescence. This switchable Myc stability probe could then be used to rapidly screen for regulators of Myc turnover. This concept has been validated and this new research tool developed, as proposed.