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

在第一阶段的资金，我们已经证明，短期葡萄糖饥饿调节Ehmeth活性通过结合烯醇化酶，糖酵解酶Ehmeth。我们将集中我们的研究肌钙蛋白，第二Dnmt 2相互作用的蛋白质，最近已确定的表征。与烯醇化酶的行为相反，在葡萄糖饥饿的寄生虫中，较少的肌钙蛋白与Ehmeth相互作用。这种相互作用的意义仍然是未知的，它将在这个财团的框架内，通过使用生物化学和分子生物学技术相结合的研究。接下来，我们将讨论其他环境因素，包括长期葡萄糖饥饿，一氧化氮和热休克，对Ehmeth的表达及其活性的影响。我们还将更详细地研究retroelements在Ehmeth表达激活中的作用。为了理解为什么Ehmeth表达在我们的实验室菌株中如此低，我们将表征其启动子区域和调控元件的存在。这种全球性的方法将有助于我们了解营养和环境压力对Dnmt 2表达的作用，这一现象也在该联盟研究的生物体中观察到。

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.