Study of chloroplast stromules during PCD and inter-organellar communication

PCD 和细胞间通讯过程中叶绿体基质的研究

• 批准号: 9382027
• 负责人: Jeffrey L Caplan
• 金额: $ 32.22万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2019-05-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9382027
• 关键词: Actins; Animals; Apoptosis; Binding; Biological Models; Carrier Proteins; Cell Death; Cell Nucleus; Cell Survival; Cell membrane; Cell surface; Cellular biology; Chloroplasts; Communicable Diseases; Communication; Complex; Computing Methodologies; Cytoplasm; Cytoskeleton; Development; Disease; Flagellin; Generations; Genetic; Goals; Growth and Development function; Hormones; Hydrogen Peroxide; Hypersensitivity; Immune; Immune response; Immune signaling; Immunity; Immunologic Receptors; Infection; Innate Immune Response; Knock-out; Leucine-Rich Repeat; MAP Kinase Gene; Malignant Neoplasms; Mammals; Mass Spectrum Analysis; Measures; Membrane; Microfilaments; Microtubules; Mitochondria; Modeling; Molecular; Morphology; Movement; NADPH Oxidase; Natural Immunity; Nuclear; Nucleotides; Organelles; Pattern; Pattern recognition receptor; Phosphotransferases; Plants; Play; Process; Proteins; Proteomics; Publishing; RIPK1 gene; Reactive Oxygen Species; Recruitment Activity; Regulation; Research; Role; Salicylic Acids; Signal Transduction; Signaling Protein; Source; Testing; Tubular formation; Vertebrates; Virulence; base; extracellular; genetic analysis; genetic approach; immune function; insight; mutant; novel; optogenetics; organelle movement; pathogen; prevent; protein biomarkers; protein transport; receptor; response; superoxide-generating NADPH oxidase; tool

Project Summary This project focuses on chloroplasts as central player in the generation of immune signals and the regulation of programmed cell death (PCD) required for innate immune responses against pathogen infection. Although mitochondria play a central role during mammalian PCD, emerging evidence suggests that in plants chloroplasts have a critical function in executing localized PCD that limits pathogen spread. Chloroplasts in addition to being involved in the generation of immune signals such as reactive oxygen species (ROS) and defense hormone salicylic acid (SA), also participate directly in the recognition of pathogen. Interestingly, the chloroplasts dynamically change their morphology during immune responses and send out tubular projections called stromules. These induced stromules use the cytoskeleton to extend and then anchor to the nucleus, which facilitate perinuclear clustering of chloroplasts and transport chloroplast-generated ROS and defense proteins to the nucleus. The overall goal of this application is to use combination of novel cell biology, genetics, proteomics and computational approaches to unravel the mechanistic basis of stromule driven periunclear chloroplast clustering and determine the identity and function of immune signals released from chloroplasts to nuclei. Specifically, Aim 1 will examine organelle and cytoskeleton dynamics during plant innate immunity. Novel computational methods quantifying stromule features and dynamics will be further developed and applied to understand chloroplasts, their stromules, and cytoskeleton dynamics during innate immunity. Both plant and animal pathogens target the cytoskeleton, and this aim will examine how pathogen effectors alter the cytoskeleton and related organelle dynamics as a virulence strategy. Aim 2 will investigate immune signals required for stromule induction and the release of chloroplast signals. The relationship of different ROS sources, organelle movement and PCD during immune responses will be examined using a combination of genetic tools and the computational methods developed in Aim 1. In Aim 3, known chloroplast-derived immune signals will be characterized and novel protein signals will be identified. The downstream targets of H2O2 in the nucleus will be elucidated and studied. Mass spectrometry will be used to identify proteins that are transported from chloroplasts to nuclei. Genetic approaches will be employed to examine their role during PCD. Lastly, the mechanism of release from chloroplasts during innate immunity will be studied to determine if it is akin to mitochondrial release during apoptosis in mammals. Understanding the role of different organelles during PCD and innate immunity will provide a unified mechanistic basis of cell death and cell survival process that occur in response to infectious pathogens. The results from our model systems will impact broadly on understanding of organelle-to-nuclear communication that influence innate immunity against infectious diseases.

项目概要 该项目重点关注叶绿体作为免疫信号生成和免疫信号生成的核心角色。 针对病原体感染的先天免疫反应所需的程序性细胞死亡（PCD）的调节。 尽管线粒体在哺乳动物 PCD 过程中发挥着核心作用，但新出现的证据表明，在植物中 叶绿体在执行限制病原体传播的局部 PCD 方面具有关键功能。叶绿体在 除了参与活性氧 (ROS) 等免疫信号的产生和 防御激素水杨酸（SA），也直接参与病原体的识别。有趣的是， 叶绿体在免疫反应过程中动态改变其形态并发出管状投射 称为斯特鲁姆斯。这些诱导的基质使用细胞骨架延伸，然后锚定到细胞核， 促进叶绿体核周聚集并运输叶绿体产生的 ROS 和防御 蛋白质进入细胞核。该应用程序的总体目标是结合使用新颖的细胞生物学、遗传学、 蛋白质组学和计算方法揭示基质驱动的核周的机制基础 叶绿体聚类并确定叶绿体释放的免疫信号的身份和功能 原子核。具体来说，目标 1 将检查植物先天免疫过程中的细胞器和细胞骨架动态。 量化基质特征和动力学的新计算方法将得到进一步开发和 用于了解先天免疫过程中的叶绿体、其基质和细胞骨架动力学。两个都 植物和动物病原体以细胞骨架为目标，这一目标将研究病原体效应子如何改变 细胞骨架和相关细胞器动力学作为毒力策略。目标 2 将研究免疫信号 基质诱导和叶绿体信号释放所需的。不同ROS的关系 将结合使用以下方法来检查免疫反应期间的来源、细胞器运动和 PCD 目标 1 中开发的遗传工具和计算方法。目标 3 中，已知叶绿体衍生的免疫 将表征信号并鉴定新的蛋白质信号。 H2O2 的下游目标 核将被阐明和研究。质谱法将用于识别运输的蛋白质 从叶绿体到细胞核。将采用遗传学方法来检查它们在 PCD 过程中的作用。最后， 将研究先天免疫期间叶绿体释放的机制，以确定它是否类似于 哺乳动物细胞凋亡过程中线粒体的释放。了解不同细胞器在 PCD 过程中的作用 而先天免疫将为细胞死亡和细胞存活过程提供统一的机制基础。 对传染性病原体的反应。我们的模型系统的结果将广泛影响对 影响针对传染病的先天免疫的细胞器到核的通讯。

项目成果

Plant-microbe interactions: organelles and the cytoskeleton in action.

植物-微生物相互作用：细胞器和细胞骨架的作用。

• DOI: 10.1111/nph.14959
• 发表时间: 2018-02-01
• 期刊: The New phytologist
• 作者: Eunsook Park;Alex Nedo;J. Caplan;S. Dinesh
• 通讯作者: S. Dinesh

Spatiotemporal Monitoring of Pseudomonas syringae Effectors via Type III Secretion Using Split Fluorescent Protein Fragments.

使用分裂荧光蛋白片段通过 III 型分泌物对丁香假单胞菌效应器进行时空监测。

• 发表时间: 2017-07
• 作者: Park, Eunsook;Lee, Hye;Woo, Jongchan;Choi, Doil;Dinesh
• 通讯作者: Dinesh

The role of TIR-NBS and TIR-X proteins in plant basal defense responses.

TIR-NBS 和 TIR-X 蛋白在植物基础防御反应中的作用。

• 发表时间: 2013-07
• 影响因子: 7.4
• 作者: Nandety, Raja Sekhar;Caplan, Jeffery L;Cavanaugh, Keri;Perroud, Bertrand;Wroblewski, Tadeusz;Michelmore, Richard W;Meyers, Blake C
• 通讯作者: Meyers, Blake C

Perspectives on intracellular perception of plant viruses.

植物病毒细胞内感知的观点。

• 发表时间: 2019
• 影响因子: 4.9
• 作者: Meier, Nathan;Hatch, Cameron;Nagalakshmi, Ugrappa;Dinesh
• 通讯作者: Dinesh

The conformational and subcellular compartmental dance of plant NLRs during viral recognition and defense signaling.

病毒识别和防御信号传导过程中植物 NLR 的构象和亚细胞区室舞蹈。

• DOI: 10.1016/j.mib.2014.05.003
• 发表时间: 2014-08
• 期刊: Current opinion in microbiology
• 影响因子: 5.4
• 作者: Padmanabhan MS;Dinesh-Kumar SP
• 通讯作者: Dinesh-Kumar SP