Ehrlichia Notch SLiM-activated oncoprotein inhibition of apoptosis

埃里希氏菌Notch SLiM激活的癌蛋白抑制细胞凋亡

基本信息

批准号：
10513824
负责人：
JERE W MCBRIDE
金额：
$ 40万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-11-01 至 2026-10-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10513824
关键词：
Apoptosis Apoptosis Inhibitor Apoptotic BCL-2 Protein BIR Domain Bacteria Binding CASP3 gene Caspase Cell Communication Cell Survival Cell physiology Cells Development Disease Down-Regulation Ehrlichia Ehrlichia chaffeensis Ehrlichiosis Etiology Eukaryotic Cell Event FBXW7 gene Genetic Transcription Genome Goals Host Defense Host Defense Mechanism Human Immune Immune Evasion Infection Inflammation Inhibition of Apoptosis Investigation Knowledge Laboratories Life Ligands Link MCL1 gene Mediating Membrane Proteins Mitochondria Molecular Molecular Mimicry Mononuclear Notch Signaling Pathway Oncoproteins Pathway interactions Phagocytes Play Positioning Attribute Post-Translational Protein Processing Protein Inhibition Proteins Receptor Signaling Regulation Research Role Science Signal Pathway Signal Transduction Surface System TLR2 gene Tandem Repeat Sequences Toll-like receptors in vivo mimetics mimicry new therapeutic target notch protein novel therapeutic intervention pathogen prevent programs protein protein interaction receptor receptor binding receptor expression tick-borne ubiquitin ligase

项目摘要

ABSTRACT Ehrlichia chaffeensis (E. ch.) is a gram-negative, obligately intracellular bacterium and the etiologic agent of human monocytotropic ehrlichiosis (HME), an emerging, life-threatening, tick-borne zoonosis. E. ch. preferentially infects mononuclear phagocytes and survives intracellularly by subverting innate immune defenses mediated in part by tandem repeat protein (TRP) effectors. Within the last decade, our laboratory has identified a multitude of molecular ehrlichial TRP-host interactions, many that are new to science, illuminating the breath and complexity of pathogen-host interaction dynamics that occur during infection. We have shown that E. ch. is dependent on activation of conserved eukaryotic signaling pathways including Notch and Wnt for infection. However, understanding the molecular basis of Notch activation events, whereby E. ch. repurposes Notch signaling for infection remains a major gap in our knowledge. Thus, the purpose of this investigation is to define the molecular and cellular mechanisms E. ch. has evolved to repurpose Notch signaling for infection. We propose that E. ch. TRP120 has distinct eukaryotic protein interaction modules known as short linear motifs (SLiMs) that mimic Notch ligand function and directly engage cognate receptors to exploit Notch signaling for infection. This investigation will address our limited understanding of the functionally diverse roles of protein interaction modules in the continuum of host-pathogen interactions and cellular reprogramming. The long-term goal of this research is to define the molecular basis of Ehrlichia host cell mimicry, and the mechanisms involved in infection and immune evasion. The objective of this proposal is to determine the molecular interactions involved and functional mechanisms whereby E. ch. TRP120 Notch ligand mimicry establishes and promotes infection by inhibiting host cell apoptosis. We hypothesize that E. ch. TRP120 has a Notch SLiM mimetic that activates Notch signaling to upregulate anti-apoptotic regulators (MCL1 and NICD), thereby inhibiting mitochondrial apoptotic signaling and caspase activation. Aim 1 will define the E. ch. TRP120 Notch SLiM mimetic and investigate receptor binding and signaling; Aim 2 will examine E. ch. TRP120 Notch-upregulated MCL1 inhibition of mitochondrial apoptosis; and Aim 3 will investigate the role of E. ch. Notch stabilization of XIAP and inhibition of caspase activation. This investigation will extend our knowledge of the molecular interactions by which E. ch. TRP120 surface protein exploits Notch signaling to inhibit apoptosis and promote host cell survival and infection. The significance of this research is defining the mechanistic strategies whereby intracellular pathogens with small genomes and a limited number of effector proteins, have evolved host mimicry modules to repurpose host cell signaling to manipulate downstream host defense mechanisms for infection. A molecular understanding of E. ch. pathobiology will also facilitate development of novel therapeutic approaches for Ehrlichia spp. and intracellular pathogens that utilize SLiM mimicry or exploit conserved cellular pathways for infection and immune evasion.

抽象的恰菲埃里希体 (E. ch.) 是一种革兰氏阴性、专性细胞内细菌，是下列疾病的病原体：人类单核埃利希体病 (HME)，一种新出现的、危及生命的蜱传人畜共患病。 E.ch. 优先感染单核吞噬细胞并通过破坏先天免疫防御在细胞内存活部分由串联重复蛋白（TRP）效应子介导。在过去的十年里，我们的实验室已经确定多种埃利希体TRP-宿主相互作用，其中许多对于科学来说是新的，阐明了呼吸以及感染期间发生的病原体-宿主相互作用动态的复杂性。我们已经证明了 E. ch。是依赖于保守的真核信号通路（包括Notch和Wnt）的激活来进行感染。然而，了解Notch激活事件的分子基础，即E. ch。重新利用缺口感染信号仍然是我们知识的一个主要空白。因此，本次调查的目的是确定分子和细胞机制 E. ch.已经进化到重新利用Notch信号来进行感染。我们建议那个E.ch。 TRP120 具有独特的真核蛋白质相互作用模块，称为短线性基序 (SLiM)，模仿Notch配体功能并直接与同源受体结合以利用Notch信号传导进行感染。这研究将解决我们对蛋白质相互作用模块的功能多样性作用的有限理解在宿主-病原体相互作用和细胞重编程的连续体中。本研究的长期目标旨在定义埃里希体宿主细胞拟态的分子基础，以及感染和感染所涉及的机制免疫逃避。该提案的目的是确定所涉及的分子相互作用和功能 E. ch 的机制。 TRP120 Notch配体拟态通过抑制宿主来建立和促进感染细胞凋亡。我们假设 E. ch。 TRP120 具有 Notch SLiM 模拟物，可激活 Notch 信号上调抗凋亡调节因子（MCL1 和 NICD），从而抑制线粒体凋亡信号传导半胱天冬酶激活。目标 1 将定义 E.ch。 TRP120 Notch SLiM 模拟物并研究受体结合和信号发送；目标 2 将检查 E. ch。 TRP120 Notch 上调 MCL1 对线粒体凋亡的抑制；目标 3 将调查 E. ch 的作用。 XIAP 的缺口稳定和半胱天冬酶激活的抑制。这研究将扩展我们对 E. ch. 分子相互作用的了解。 TRP120表面蛋白利用 Notch 信号传导抑制细胞凋亡并促进宿主细胞存活和感染。此举的意义研究正在定义具有小基因组和有限的细胞内病原体的机制策略许多效应蛋白，已进化出宿主拟态模块，以重新利用宿主细胞信号传导来操纵感染的下游宿主防御机制。对 E. ch 的分子理解。病理学也将促进埃里希体属新治疗方法的开发。和利用的细胞内病原体 SLiM 模仿或利用保守的细胞途径进行感染和免疫逃避。