Biophysical Characterization of the Yorkie-Mask Complex Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/d217qr500

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  • Yes-associated protein (YAP) in mammals and Yorkie (Yki) in Drosophila are functionally homologous proteins that partner with transcription factors to promote cell proliferation. The Drosophila protein Mask and its mammalian homolog MASK1 are examples of transcription factors that are essential for Yki/ YAP to drive transcription of target genes. Increased Mask activity increases Yki expression; overexpression of Yki is associated with cancer and tumor formation. Yki is reported to bind three domains of Mask in Drosophila S2 cells, but it is not clear exactly where Mask binds Yki. In this in vitro study, we overexpressed, purified and characterized the solution properties of Yki, and two of the three putative binding domains of Mask. Using chemical crosslinking, native gel shift assay, and pull-down assay experiments, we did not detect binding between our recombinant Yki and the two recombinant Mask domains. Future directions will focus on interactions between Yki, and the third putative binding domain of Mask.
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