The Function of EBV LMP1 CTAR3 in Sumoylation and Oncogenesis

EBV LMP1 CTAR3 在 Sumoylation 和肿瘤发生中的功能

• 批准号: 9055658
• 负责人: Gretchen L Bentz
• 金额: $ 24.62万
• 依托单位: MERCER UNIVERSITY MACON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-20 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9055658
• 关键词: Affect; Amino Acids; Animal Model; Award; Behavior; Biological Models; Biological Process; Biology; C-terminal; Cell Line; Cell Nucleus; Cell Survival; Cells; Cytoskeletal Proteins; DNA-Protein Interaction; Data; Disease; Environment; Enzymes; Epithelial; Epstein-Barr Virus Infections; Epstein-Barr pathogenesis; Epstein-Barr virus LMP-1 protein; Event; Faculty; Goals; Human Herpesvirus 4; Human Herpesvirus 8; Immune response; Interferon Activation; Knock-out; Knowledge; Learning; Lymphoid; Lytic; Lytic Phase; Malignant Neoplasms; Mediating; Membrane; Membrane Proteins; Mentors; Mus; Nucleic Acid Regulatory Sequences; Oncogenes; Oncogenic; Pathology; Pathway interactions; Peptide Hydrolases; Phase; Phosphorylation; Post-Translational Protein Processing; Process; Proteins; Publications; Publishing; Regulation; Reporting; Repression; Research; Research Personnel; Response Elements; Role; STAT1 gene; Scientist; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Techniques; Technology; Testing; Training; Ubiquitination; Viral; Virus; Work; Zinc Fingers; abstracting; career; cell motility; cellular targeting; gammaherpesvirus; humanized mouse; immunogenic; in vivo; insight; latent infection; lytic replication; member; mouse model; protein function; protein protein interaction; skills; trafficking; transcription factor; tumor; tumorigenesis; tumorigenic

Project Abstract: Candidate and Environment: My career goal is to become a leading independent scientist in the field of herpesvirology. During the mentored phase of the Pathway to Independence Award, I will work to further establish my publication record in the field of EBV, laying the groundwork for the research I will do as a faculty member. This project will provide training in new techniques [BAC technology, a humanized mouse model system, investigating protein-protein and protein-DNA interactions] that will be applicable to the research performed as an independent investigator. The expertise of Dr. Pagano as well as other faculty members at UNC will be invaluable in learning the necessary skills to become a successful independent researcher. Research: EBV, a ubiquitous γ-herpesvirus, is associated with several lymphoid and epithelial malignancies. Latent membrane protein-1 (LMP1), the principal viral oncoprotein, is a constitutively active membrane signaling protein that regulates multiple signal transduction pathways via its cytoplasmic C-terminal activating regions (CTAR). In contrast to CTAR1 and CTAR2, the function for CTAR3 is not well defined. I recently published that LMP1 CTAR3 is necessary and sufficient for LMP1 to interact with the SUMO-conjugating enzyme Ubc9 and induce the sumoylation of cellular proteins. Disruption of the LMP1-Ubc9 interaction resulted in changes in cellular behaviors associated with oncogenesis and LMP1 and loss of LMP1-induced sumoylation. Further understanding how LMP1 CTAR3 functions and mechanisms by which it functions is my next priority. I hypothesize that CTAR3 is a self-regulatory domain that helps modulate the oncogenic potential of LMP1. In Specific Aim 1, to be completed during the mentored phase, I will determine biological functions of LMP1 CTAR3 in vivo. Using wildtype and LMP1 CTAR3 deletion viruses to infect humanized mice, a new animal model developed at LCCC for EBV infection and disease, I will examine the tumorigenic and immunogenic potentials of the viruses. The role of CTAR3 is the intracellular trafficking and stability of LMP1 will also be tested. In Specific Aim 2, to be completed during the independent phase, I will examine mechanisms by which CTAR3 functions. Initial targets of LMP1-induced sumoylation include IRF7, STAT1, and the SUMO protease SENP1. I will investigate the role of LMP1 CTAR3 in the sumoylation of these proteins and how sumoylation regulates protein function. Finally, because protein sumoylation can help inhibit the latent-lytic switch, I will examine in LMP1 CTAR3 contributes to LMP1-mediated repression of lytic replication by inducing protein sumoylation, specifically of PML, ZEB1, and ZEB2. The proposed work will greatly increase our understanding of functions for LMP1 CTAR3, specifically its role in regulating protein sumoylation, and how CTAR3 functions, expanding knowledge of how LMP1 acts as an oncoprotein.

项目摘要： 候选人和环境：我的职业目标是成为一个领先的独立科学家在该领域的 疱疹病毒学在独立之路奖的指导阶段，我将努力进一步 建立我在EBV领域的出版记录，为我作为一名教师所做的研究奠定基础 会员是否这个项目将提供新技术的培训[生物活性炭技术，一个人源化的小鼠模型 系统，研究蛋白质-蛋白质和蛋白质-DNA相互作用]，将适用于研究 作为一名独立调查员。帕加诺博士以及其他教职员工的专业知识， 这将是宝贵的学习必要的技能，成为一个成功的独立研究人员。 研究：EBV是一种普遍存在的γ-疱疹病毒，与几种淋巴和上皮恶性肿瘤有关。 潜伏膜蛋白1（LMP 1）是病毒的主要癌蛋白，是一种组成型活性膜 一种信号蛋白，通过其胞质C-末端激活调节多种信号转导途径 区域（CTAR）。与CTAR 1和CTAR 2相反，CTAR 3的功能没有很好的定义。我最近 LMP 1CTAR 3是LMP 1与SUMO结合蛋白相互作用所必需和充分的。 酶Ubc 9并诱导细胞蛋白质的类小泛素化。LMP 1-Ubc 9相互作用的破坏导致 与肿瘤发生和LMP 1相关的细胞行为变化以及LMP 1诱导的细胞凋亡的丧失 类小泛素化进一步了解LMP 1 CTAR 3的功能和机制，它的功能是我 下一个优先事项我假设CTAR 3是一个自我调节结构域，有助于调节致癌基因， LMP 1的潜力在具体目标1中，将在指导阶段完成，我将确定生物学 LMP 1 CTAR 3在体内的功能。使用野生型和LMP 1 CTAR 3缺失病毒感染人源化小鼠， 一种新的动物模型，在LCCC开发的EBV感染和疾病，我将检查致瘤性和 病毒的免疫原性潜力。CTAR 3的作用是LMP 1的细胞内运输和稳定性 也将受到考验。在独立阶段完成的具体目标2中，我将研究 CTAR 3发挥作用的机制。LMP 1诱导的类小泛素化的初始靶点包括IRF 7、STAT 1和 SUMO蛋白酶SENP 1。我将研究LMP 1 CTAR 3在这些蛋白质类小泛素化中的作用， 类小泛素化如何调节蛋白质功能。最后，由于蛋白质类小泛素化可以帮助抑制潜在的溶解， 开关，我将检查在LMP 1 CTAR 3有助于LMP 1介导的抑制裂解复制诱导 蛋白类小泛素化，特别是PML，ZEB 1和ZEB 2。拟议的工作将大大增加我们的 了解LMP 1 CTAR 3的功能，特别是它在调节蛋白质类小泛素化中的作用，以及如何 CTAR 3功能，扩展了LMP 1如何作为癌蛋白的知识。