Structural and Functional Characterization of METTL13

METTL13 的结构和功能表征

• 批准号: 10538038
• 负责人: Krystal Diaz
• 金额: $ 4.68万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-11 至 2023-08-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10538038
• 关键词: Adherence; Amino Acid Motifs; Amino Acids; Apoptosis; Binding; Biological; Biological Assay; C-terminal; CRISPR/Cas technology; Cancer cell line; Catalysis; Cell Line; Cell Proliferation; Cell physiology; Chromatin; Consensus; Crystallization; DNA Repair; Defect; Development; Disease; Docking; Enzymes; Event; Functional disorder; Future; Generations; Goals; Human; Knock-out; Link; Literature; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Malignant neoplasm of urinary bladder; Mediating; Methylation; Methyltransferase; Mitosis; Molecular; N-terminal; NMR Spectroscopy; Outcome; Output; Pathogenesis; Pathway interactions; Peptide Elongation Factor 1; Peptides; Play; Post-Translational Protein Processing; Prognosis; Proteins; Regulation; Reporting; Research; Role; S-Adenosylmethionine; Site; Structure; Substrate Specificity; Testing; Thermodynamics; Tissue Model; Tissues; Translations; Tumor Suppression; Up-Regulation; base; bladder Carcinoma; cancer cell; cofactor; functional outcomes; in vivo; insight; migration; mutant; novel; overexpression; targeted cancer therapy; tumor progression; tumorigenesis

PROJECT SUMMARY Advances in research on protein α-N-terminal (Nα) methylation have brought to light the important role of this post-translational modification in the regulation of mitosis, chromatin interactions, and DNA repair. Altered methylation levels result in the aberrant expression of N-terminal methyltransferases (NTMTs); this aberrant expression is often implicated in the development of various diseases. However, the function of N-terminal methylation and its role in disease pathogenesis remains poorly characterized. Until recently, N-terminal methylation was believed to only occur on a conserved X-P-K motif of protein substrates and catalyzed by only two enzymes, NTMT1 and NTMT2. Identification of N-terminal methylation on the eukaryotic elongation factor 1 alpha (eEF1A) GKEK motif, a non-canonical NTMT motif, has challenged this paradigm, bringing to light the possibility for additional existing NTMT substrates. The methyltransferase like 13 (METTL13) enzyme has very recently been identified as the methyltransferase responsible for eEF1A N-terminal methylation. METTL13 is a dual methyltransferase that can methylate both the N-terminus of Lys55 of eEF1A through its separate methyltransferase domains, but the biological roles of these methylations are currently unclear. The implication of both METTL13 and eEF1A in the poor prognosis associated with pancreatic, lung, and bladder cancers motivates us to interrogate the role of this novel methylation event in cellular dysfunction. The downstream effects of dysfunctional METTL13 substrate recognition and catalysis and their relation to cancer development are entirely unknown, which highlights a critical need to identify the structural and mechanistic details of METTL13- mediated Nα methylation that govern cancer pathogenesis and progression. My central hypothesis is that METTL13-mediated eEF1A Nα methylation may serve a distinct functional role from eEF1A Lys55 methylation and their dysregulation may yield different biological outcomes in a tissue-specific manner. The long-term goals of this proposal are to (1) generate a comprehensive understanding of the substrate recognition, regulation, and function of Nα methylation catalyzed by METTL13 and (2) determine the relation between METTL13-mediated Nα methylation and cancer progression. The results from this research will clarify the biological role of Nα methylation and define structural features of METTL13 that have the potential to inform future targeted cancer therapies.

项目摘要 蛋白质α-N-末端甲基化的研究进展揭示了其在蛋白质合成中的重要作用 这种翻译后修饰在有丝分裂、染色质相互作用和DNA修复的调节中起着重要作用。改变 甲基化水平导致N-末端甲基转移酶（NTMT）的异常表达;这种异常的甲基化水平导致N-末端甲基转移酶（NTMT）的异常表达。 其表达通常与各种疾病的发展有关。然而，N-末端的功能 甲基化及其在疾病发病机制中的作用仍然缺乏表征。直到最近，N端 据信甲基化仅发生在蛋白质底物的保守X-P-K基序上，并且仅由 两种酶，NTMT 1和NTMT 2。真核细胞延伸因子1 N端甲基化的鉴定 α（eEF 1A）GKEK基序，一种非经典的NTMT基序，挑战了这种范式，揭示了 增加现有NTMT底物的可能性。甲基转移酶样13（Methyltransferase like 13）酶 最近被鉴定为负责eEF 1A N-末端甲基化的甲基转移酶。L13是一种 双甲基转移酶，其可以通过其单独的甲基化eEF 1A的Lys 55的N-末端 甲基转移酶结构域，但这些甲基化的生物学作用目前尚不清楚。言下之意 与胰腺癌、肺癌和膀胱癌相关的不良预后中的胃L13和eEF 1A 促使我们探究这种新的甲基化事件在细胞功能障碍中的作用。下游效应 功能失调的胃L13底物识别和催化及其与癌症发展的关系， 完全未知，这突出了确定胃L13的结构和机制细节的迫切需要- 介导的Nα甲基化控制癌症的发病机制和进展。我的核心假设是 EPL 13介导的eEF 1A Nα甲基化可能与eEF 1A Lys 55甲基化具有不同的功能作用 并且它们的失调可能以组织特异性方式产生不同的生物学结果。远景目标 该提案的目的是（1）全面了解底物识别，监管， （2）确定胃L13介导的Nα甲基化与胃蛋白酶活性之间的关系。 Nα甲基化与癌症进展这项研究的结果将阐明Nα的生物学作用 甲基化和定义有可能告知未来靶向癌症的胃L13的结构特征 治疗