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 本应用的总体目标是阐明Lys63(K63)连接的多泛素化的机制。 细胞外信号调节激酶ERK1和ERK2，并确定这一新的翻译后作用 对肿瘤发生的修饰。ERK1和ERK2具有较高的结构和功能 相似的是有丝分裂原激活的蛋白激酶(MAPK)级联的下游效应器 细胞行为和细胞命运决定对广泛的细胞内信号的反应。ERK信号转导 有丝分裂途径是哺乳动物细胞的主要有丝分裂途径。它在很大一部分中异常上调 由于上游信号成分突变和ERK重新激活所致的人类肿瘤 也是针对这些上游成分的药物产生耐药性的最常见机制。发展中 一种适用于由过度活跃的ERK通路驱动的广泛肿瘤的疗法，并且可以 规避耐药性仍然是癌症研究中的一大挑战。应对这一挑战 需要全面了解ERK是如何在MAPK中被具体有效地激活的 卡斯卡德。目前对ERK激活的了解很大程度上局限于其上游的磷酸化。 激酶MEK。在我们的初步研究中，我们发现ERK与K63连接的多泛素链连接， 这种翻译后修饰与ERK的激活有关。此外，我们已经确定了 三段基序蛋白TRIM15作为泛素连接酶，肿瘤抑制因子CyLD作为去泛素化 酶(DUB)，可动态调节ERK泛素化。在这里，我们将检验中心假设 K63泛素化ERK对于这些激酶的特异性和高效激活以及这种失调是至关重要的 这种翻译后修饰与肿瘤的发病机制和治疗耐药有关。我们 提出三个具体目标。首先，我们将研究TRIM15和CyLD在K63泛素化过程中的作用。 ERK，并确定它们与ERK的相互作用如何受到有丝分裂刺激的调节。第二，我们将澄清 K63泛素化在ERK激活中的作用和机制。三是明确党的作用 TRIM15和CyLD与肿瘤发生和耐药的关系总体而言，这些目标将解决 ERK生物学和致癌信号转导的基本问题，并将为 针对过度活跃的ERK驱动的人类肿瘤的有效治疗方法的发展 信号通路。