Identification of Cytoplasmic Proteins Interacting with the Mammary Cell Transforming Domain of Ese-1
Annual rept. 29 Mar 2006-28 Mar 2007
COLORADO UNIV HEALTH SCIENCES CENTER AURORA CO
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The Ets family of transcription factors contains several members that are important components of the cellular pathways leading to tumorigenesis. The Ese-1 gene is an Ets member that is particularly relevant to breast cancer. Ese-1 is located on chromosome 1q32.1, in a region that is amplified in 50 of early breast cancers. Ese-1 mRNA is over-expressed in human breast ductal carcinoma in situ DCIS. We recently discovered that Ese-1 transforms MCF-12A cells via a novel cytoplasmic mechanism in which a unique 40-amino acid AA, serine- and aspartic acid rich SAR domain is necessary and sufficient for transformation. Furthermore, we reported that Ese-1 protein is abundantly expressed in the cytoplasm of human ductal carcinoma in situ DCIS specimens. However, identification of the cytoplasmic partners of the Ese-1 SAR motif and the precise mechanism by which cytoplasmic signaling mediated by the Ese-1 SAR motif occurs remain unknown. In this application, we propose to take advantage of this novel paradigm. Specifically, we plan to apply innovative and powerful state-of-the-art molecular yeast two-hybrid and protein biophysical methods MALDI-TOF and LC-mass spectrometry to identify proteins that interact with the Ese-1 SAR transformation domain. We will then validate which partners of Ese-1 are functionally relevant to breast cancer. We have made diligent progress, having identified a putative interactor, which we are validating.
- Medicine and Medical Research