Regulation of NRMT1 through homolog binding

项目总结： N-末端α-氨基(N-α)的翻译后修饰是一种高度保守和广泛应用的方法 这一过程见于从细菌到哺乳动物的所有物种。Schaner Toey博士开创了N-α领域-- 通过鉴定前两个真核Nα甲基转移酶NRMT1和NRMT2(也称为 METTL11A和METTL11B)。她接着描述了这些酶的共同序列， 鉴定了它们的几十种底物，并表明N-α甲基化调节蛋白质/dna相互作用。她的实验室是 也是第一个表明，像组蛋白PTM一样，N个α-PTM是功能代码的一部分。他们确定了第一个 已知的蛋白质以Nα-乙酰和Nα-甲基两种形式存在，肌球蛋白轻链9(MYL9)。他们 证明这两个Nα-PTM产生不同的MYL9蛋白形式，具有独特的蛋白质结合 伙伴、内部PTM模式和细胞隔间特有的功能。他们还展示了 NRMT1在肿瘤发生和衰老中的重要作用。乳腺癌细胞中NRMT1的缺失促进 增殖、迁移、克隆形成和包裹体生长。小鼠NRMT1基因敲除促进 与过早衰老有关的表型，包括早期白发和脱发、脊柱后凸、皮肤和肝脏 纤维化、神经变性、记忆力丧失和寿命缩短。他们已经证明了NRMT1调节 神经和肌肉干细胞的发展，并假设NRMT1在干细胞中的一般作用 分化导致了许多过早衰老的表型。他们现在想要1。)扩大了解 NRMT1调控的生物化学，2)鉴定由N个α-Ptm编码调控的附加底物， 和3.)从机制上描述新发现的NRMT1在干细胞命运决定中的作用。 本多样性补编中提议的工作将涉及第一点，并审查对 NRMT1通过与其METTL家族同系物、NRMT2和METTL13结合。当它变得越来越明显时 甲基转移酶调节的一种常见方法是通过形成具有紧密同源物的络合物， NRMT1是已知的第一个受两种不同同系物相反调控的甲基转移酶。在这里我们 将研究这些相互作用的生物化学和生物学相关性。成功完成这些任务 实验不仅将进一步发展NRMT1作为人类潜在的治疗靶点 癌症和与年龄相关的疾病，而且还完成了Haley Parker在技术和 继续药物设计研究生涯所需的专业技能。

Project Summary: Post-translational modification of N-terminal α-amino (Nα) groups is a highly conserved and vastly utilized process seen in all species from bacteria to mammals. Dr. Schaner Tooley pioneered the field of Nα- methylation by identifying the first two eukaryotic Nα-methyltransferases, NRMT1 and NRMT2 (also known as METTL11A and METTL11B). She went on to characterize the consensus sequence of these enzymes, identify dozens of their substrates, and show Nα-methylation regulates protein/DNA interactions. Her lab was also the first to show that, like histone PTMs, Nα-PTMs are part of a functional code. They identified the first protein known to exist in both Nα-acetyl and Nα-methyl forms, Myosin Light Chain 9 (MYL9). They demonstrated that these two Nα-PTMs create distinct proteoforms of MYL9, with unique protein binding partners, internal PTM patterns, and cell compartment-specific functions. They have also demonstrated important roles for NRMT1 in oncogenesis and aging. NRMT1 loss in breast cancer cells promotes proliferation, migration, colony formation, and xenograph growth. NRMT1 knockout in mice promotes phenotypes associated with premature aging, including, early graying and hair loss, kyphosis, skin and liver fibrosis, neurodegeneration, memory loss, and decreased lifespan. They have shown that NRMT1 regulates both neural and muscle stem cell development, and hypothesize a general role for NRMT1 in stem cell differentiation drives many of the premature aging phenotypes. They now want to 1.) expand understanding of the biochemistry of NRMT1 regulation, 2.) identify additional substrates regulated by the Nα-PTM code, and 3.) mechanistically characterize the newly discovered role for NRMT1 in stem cell fate determination. The worked proposed in this diversity supplement will address the first point and examine the regulation of NRMT1 through binding of its METTL family homologs, NRMT2 and METTL13. While it is becoming apparent that a common method of methyltransferase regulation is through complex formation with close homologs, NRMT1 is the first methyltransferase known to be opposingly regulated by two different homologs. Here we will study both the biochemistry and biological relevance of these interactions. Successful completion of these experiments will not only further the development of NRMT1 as a potential therapeutic target for human cancers and age-related disorders, but also complete Haley Parker's training in the technical and professional skills necessary to continue a research career in drug design.