Characterization of E.histolytica DNMT2 homolog EhDNMT2 and the meaning of its interaction with Enolase

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During the first period of funding, we have demonstrated that short term glucose starvation regulates Ehmeth activity via the binding of enolase, a glycolytic enzyme to Ehmeth. We will concentrate our research on the characterization of troponin, a second Dnmt2 interacting protein that has been recently identified. In contrast to the behavior of enolase, less troponin interacts with Ehmeth in glucose starved parasites. The meaning of this interaction is still unknown and it will be studied within the framework of this consortium by using a combination of biochemical and molecular biology techniques. Next, we would like to address the influence of other environmental factors including long term glucose starvation, nitric oxide and heat shock, on the expression of Ehmeth and on its activity. We will also investigate more in details the role of retroelements in the activation of Ehmeth expression. In an effort to understand why Ehmeth expression is so low in our laboratory strain, we will characterize its promoter region and the presence of regulatory elements. This global approach will help us to understand the role of nutritional and environmental stresses on Dnmt2 expression, a phenomenon that has also been observed among the organisms studied in this consortium.