Interrogating Functions of Protein Methyltransferases with Chemical Tools

项目摘要/摘要：蛋白质组范围的蛋白质甲基化是一种表观遗传事件， 受&gt；60蛋白甲基转移酶(PMT；60)调节。因为传统的方法只能 在定义的细胞中发现一小部分PMT底物而不知道PMT甲基组 背景下，我们对PMT在许多基本生物学中的表观遗传学作用(S)只有部分了解 设置。对PMT的选择性扰动对于阐明它们的作用也很有用(S)。而PMT- 相关生物学通常是通过基因扰动来探测的，这种方法不同于小分子 抑制剂，缺乏时间(开启/关闭)、空间(特定部位)和剂量(一系列疗效)控制。 此外，PMTs的遗传和药物干扰的结果可能是不同的。 因为前者完全消融PMT蛋白，而后者只影响一部分 PMT的功能。然而，开发选择性和有效的PMTs小分子抑制剂是 很有挑战性。这项提议的目标是整合新的化学工具(高质量 抑制剂和PMTs的生物正交性探针)与常规方法检测 PMT的药理抑制的结果(S)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处理它们，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.