Regulation of the ERK signaling pathway by K63-linked polyubiquitination

K63 连接的多聚泛素化对 ERK 信号通路的调节

• 批准号: 10535249
• 负责人: Xiaolu Yang
• 金额: $ 40.46万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-09 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10535249
• 关键词: Address; Amino Acid Motifs; BRAF gene; Biochemical; Biology; Cell Line; Cells; Deubiquitinating Enzyme; Development; Drug Targeting; Drug resistance; Etiology; Eukaryota; Extracellular Signal Regulated Kinases; FDA approved; Goals; Guanosine Triphosphate Phosphohydrolases; Human; Hyperactivity; Knowledge; Light; Link; MAP Kinase Kinase Kinase; MAP3K1 gene; MAPK1 gene; MAPK3 gene; MEKs; Malignant Neoplasms; Mammalian Cell; Melanoma Cell; Metastatic Melanoma; Mitogen-Activated Protein Kinase Kinases; Mitogen-Activated Protein Kinases; Modification; Molecular; Mutation; Normal Cell; Oncogenic; Pathogenesis; Pathologic Processes; Pathway interactions; Patients; Phosphorylation; Phosphotransferases; Physiological Processes; Polyubiquitin; Polyubiquitination; Post-Translational Protein Processing; Prevalence; Proteins; Receptor Protein-Tyrosine Kinases; Regulation; Resistance; Role; Sampling; Signal Pathway; Signal Transduction; Site; Specific qualifier value; Specificity; Stimulus; TRIM Motif; Testing; Tumor Suppressor Proteins; Ubiquitination; Unresectable; Work; Xenograft procedure; anticancer research; base; cancer cell; cancer therapy; cell behavior; effective therapy; extracellular; inhibitor; insight; melanoma; novel; overexpression; patient derived xenograft model; protein function; prototype; resistance mechanism; response; therapy resistant; tumor; tumorigenesis; ubiquitin ligase; upstream kinase; virtual

PROJECT SUMMARY/ABSTRACT The overall goal of this application is to elucidate the mechanism of Lys63 (K63)-linked polyubiquitination of extracellular signal-regulated kinases ERK1 and ERK2, and to determine the contribution of this novel post-translational modification to tumorigenesis. ERK1 and ERK2, which share high structural and functional similarities, are downstream effectors of a mitogen-activated protein kinase (MAPK) cascade that dictates cellular behavior and cell fate decisions in response to a wide range of intracellular signals. The ERK signaling pathway is the primary mitogenic pathway in mammalian cells. It is aberrantly upregulated in a large fraction of human tumors due to prevalence of mutations in the upstream signaling components, and reactivation of ERK is also the most common mechanism for resistance to drugs that target these upstream components. Developing a therapy that is applicable for the wide range of tumors driven by a hyperactive ERK pathway and that can circumvent drug resistance remains a major challenge in cancer research. To contend with this challenge requires a comprehensive understanding of how ERK is specifically and efficiently activated within the MAPK cascade. The current knowledge of ERK activation is largely limited to their phosphorylation by the upstream kinase MEK. In our preliminary studies, we have found that ERK is conjugated to K63-linked polyubiquitin chains, and that this post-translational modification correlates with ERK activation. Furthermore, we have identified the tripartite-motif protein TRIM15 as a ubiquitin ligase, and the tumor suppressor CYLD as a deubiquitinating enzyme (DUB), that may dynamically regulate ERK ubiquitination. Here we will test the central hypothesis that K63 ubiquitination of ERK is critical for the specific and efficient activation of these kinases and that dysregulation of this post-translational modification contributes to the pathogenesis and therapeutic resistance of tumors. We propose three specific aims. First, we will characterize the role of TRIM15 and CYLD in K63 ubiquitination of ERK and define how their interactions with ERK are regulated by mitogenic stimuli. Second, we will elucidate the function and mechanism of K63 ubiquitination in the activation of ERK. Third, we will determine the role of TRIM15 and CYLD in tumorigenesis and therapeutic resistance. Collectively, these aims will address fundamental issues in ERK biology and oncogenic signaling, and will provide valuable information for the development of effective therapies for the plethora of human tumors that are driven by a hyperactive ERK signaling pathway.

项目摘要/摘要 该应用的总体目标是阐明Lys63（K63）连接的多泛素化的机理 细胞外信号调节的激酶ERK1和ERK2，并确定这种新型翻译后的贡献 修饰肿瘤发生。 ERK1和ERK2，具有较高的结构和功能 相似性是促有丝分裂原激活蛋白激酶（MAPK）级联反应的下游效应子 细胞行为和细胞命运决定响应广泛的细胞内信号。 ERK信号 途径是哺乳动物细胞中的主要有丝分裂途径。它在很大一部分中异常上调 由于上游信号成分中突变的流行和ERK重新激活而引起的人类肿瘤 也是针对这些上游成分的药物抗性的最常见机制。发展 一种适用于多活跃途径驱动的广泛肿瘤的疗法，可以 抗药性抗药性仍然是癌症研究中的主要挑战。应对这个挑战 需要全面了解ERK在MAPK中如何有效地激活 级联。当前对ERK激活的知识在很大程度上仅限于上游的磷酸化 激酶mek。在我们的初步研究中，我们发现ERK与K63连接的多泛素链相结合，， 这种翻译后修饰与ERK激活相关。此外，我们已经确定了 三方 - 米特蛋白TRIM15作为泛素连接酶，肿瘤抑制剂CyLd作为去泛素化 酶（DUB），可能会动态调节ERK泛素化。在这里，我们将测试中心假设 ERK的K63泛素化对于这些激酶的特定和有效激活至关重要 这种翻译后修饰有助于肿瘤的发病机理和治疗性。我们 提出三个具体目标。首先，我们将表征TRIM15和CYLD在K63泛素化中的作用 ERK并定义其与ERK的相互作用如何受促丝分裂刺激的调节。其次，我们将阐明 K63泛素化在ERK激活中的功能和机制。第三，我们将确定 TRIM15和CYLD在肿瘤发生和治疗性抗性中。总的来说，这些目标将解决 ERK生物学和致癌信号传导中的基本问题，并将为 开发由过度活跃的ERK驱动的过多人类肿瘤的有效疗法 信号通路。