Cloning, Purification, and Inhibition of the SUV39H2 Histone Methyl Transferase

Abstract

The most significant challenge in treating human colorectal cancer (CRC) is metastasis of the cancer. Although the study of colorectal cancer metastasis is still elementary, it is known that silencing of Fas expression is a hallmark of human colorectal cancer metastasis. The Fas protein is a member of the tumor necrosis factor (TNF)- a receptor superfamily that plays a major role in the regulation of programmed cell death. The interaction of Fas with its ligand, Fas L, triggers a signal cascade that ends in apoptosis. Silencing of Fas expression thus allows cancer cells to evade cell death triggered by the immune system. In human CRC patients, the Fas promoter is characterized by high H3K9 trimethylation (H3K9me3). This methylation is primarily caused by histone methyl transferases such as SUV39H2. We hypothesize that inhibiting H3K9 trimethylation is a potential anti-cancer strategy. To study this possibility, the gene for human SUV39H2 was cloned into pET21 and the protein was overexpressed in E.coli BL21- DE3 cells. The activity of the purified protein will be tested against various inhibitors. Funding Source: National Science Foundation