• Expression of Suv39h1 in Escherichia coli

      Baumert, Delphine; Department of Chemistry and Physics (2016-03)
      A hallmark of metastatic human colorectal cancer is silencing of Fas expression, a death receptor protein involved in an extrinsic apoptotic pathway. A number of histone methyltransferases (HMTases), including SUV39H1, have recently been linked to silencing Fas expression through methylation of histone H3 lysine 9 (H3K9) in the Fas promoter region; methylation being an epigenetic factor commonly associated with gene silencing. A natural compound found in pathogen-infected mushrooms, verticillin A, has been identified as a potent inhibitor of SUV39H1, as well as a number of other HMTases. Unfortunately it exhibits toxicity at low concentrations in mice, likely because of its association of multiple targets. Because of this toxicity and the lack of specificity exhibited by verticillin A, the goal of this project is to to identify SUV39H1-specific small molecule inhibitors. Once identified, these inhibitors will be tested for their ability to suppress colon carcinoma growth. In order to obtain large amounts of SUV39H1 for this study, we are working on sub-cloning the gene for SUV39H1 into the expression vector, pET21c(+), so that the gene product can be over-expressed in E. coli BL21 cells. Once the protein has been purified, we will test the effects of various inhibitors on the activity of SUV39H1 in vitro. Funding Source: Center for Undergraduate Research and Scholarship and Department of Chemistry and Physics