Interrogating Functions of Protein Methyltransferases with Chemical Tools

项目概要/摘要：蛋白质组范围内的蛋白质甲基化是一种表观遗传事件， 由>60种蛋白质甲基转移酶（PMT）调节。因为传统的方法只能 在不知道PMT甲基化组的情况下，在定义的细胞内暴露一小部分PMT底物， 在许多重要的生物学背景下，我们对PMT的表观遗传作用只有部分的了解。 设置.光电倍增管的选择性扰动也很有助于阐明它们的作用。虽然PMT- 相关生物学通常通过遗传扰动来探测，这种方法与小分子生物学不同， 抑制剂，缺乏时间（打开/关闭），空间（位点特异性）和剂量（一系列功效）控制。 此外，PMT的遗传干扰与药理干扰的结果可能不同 因为前者完全消除PMT蛋白，而后者仅影响PMT蛋白的子集。 PMT的功能。然而，开发选择性和有效的PMT小分子抑制剂是非常困难的。 挑战性该提案的目标是整合新型化学工具（高质量 抑制剂和PMT的生物正交活性探针）与常规方法来检查 PMT药理学抑制的结果和治疗潜力。我们计划定义 甲基化事件对基因组复制和DNA损伤反应至关重要， 以时间和剂量依赖的方式操纵它们，并使用 准确的动物模型。

Project Summary/Abstract: Proteome-wide protein methylation is an epigenetic event that is regulated by >60 protein methyltransferases (PMTs). Because conventional approaches can only uncover a small fraction of PMT substrates without knowing PMT methylome under defined cellular settings, we have only partial insight into epigenetic role(s) of PMT in many essential biological settings. Selective perturbation of PMTs is also of great use to elucidate their role(s). While PMT- associated biology is often probed via genetic perturbation, this approach, unlike small-molecule inhibitors, lacks temporal (turn on/off), spatial (loci-specific) and dose (a range of efficacy) controls. In addition, the outcomes of genetic versus pharmacological perturbation of PMTs can be different because the former completely ablates a PMT protein whereas the latter only affects a subset of PMT's functions. However, developing selective and potent small-molecule inhibitors of PMTs is challenging. The objective of this proposal is to integrate novel chemical tools (high-quality inhibitors and bioorthogonal activity probes of PMTs) with conventional methods to examine the outcome(s) and therapeutic potential of pharmacological inhibition of a PMT. We plan to define the methylation events critical for genome replication and DNA damage responses, pharmacologically manipulate them in a temporal and dose-dependent manner, and predict its outcomes using accurate animal models.