Modulation of the Effector-Triggered Immunity response by ERAD-Mediated Degradation of Activated NLRs (EMDAN)

植物抗病反应可由核苷酸结合的富含亮氨酸的重复蛋白（称为抗性蛋白）和特定病原体衍生蛋白（称为效应蛋白）的相互作用触发。这种相互作用导致强烈的防御反应，称为效应触发免疫（ETI），通常导致病原体感染部位的快速局部细胞死亡反应。这种反应必须严格调节，以产生抗性，但不能过度，以免不必要地抑制植物生长。我们的初步证据表明，玉米中常见的蛋白质降解途径的一个组成部分，ZmCER9，在激活后控制核苷酸结合的富含亮氨酸的重复蛋白的降解。ZmCER9蛋白序列与参与内质网相关降解（ERAD）过程的其他蛋白质相似，该过程可降解折叠错误的蛋白质。虽然ERAD已经在酵母中被表征，但在植物中却知之甚少。因此，ZmCER9的相互作用和随后核苷酸结合的富含亮氨酸的重复蛋白的降解似乎是先前未描述的机制，介导了激活后效应物触发的免疫防御反应的失活。我们将使用一系列技术来深入了解CER9同源物和核苷酸结合的富含亮氨酸的重复蛋白如何相互作用，控制植物的关键防御反应，并识别植物中参与该过程的其他蛋白质。这项研究将有助于更好地理解植物在面对生物压力时如何生存和茁壮成长。该项目还将为来自东北低收入农村地区的初高中学生提供培训机会和丰富STEM知识，帮助培养下一代植物科学家。植物抗病（R-）核苷酸结合富亮氨酸重复（NLR）型蛋白被激活，并在识别特定病原体衍生的效应蛋白时诱导强烈的防御反应，称为效应触发免疫或ETI。ETI的有效性取决于同源病原体缺失时的不活性，病原体识别时的快速诱导以及诱导后的快速抑制。泛素-蛋白酶体途径是所有真核生物中存在的主要蛋白质修饰和降解途径，由E1（泛素激活）、E2（泛素结合）和E3（泛素连接酶）的顺序作用介导。我们的初步数据表明，玉米e3连接酶ZmCER9介导Rp1-D NLR - r蛋白和其他几种NLR - r蛋白的降解，特别是在激活后。基于同源性，ZmCER9可能是内质网相关降解（ERAD）的一个组成部分，ERAD是一种基本的真核质量控制系统，可以降解折叠错误的蛋白质。植物ERAD的研究很少，CER9复合物的底物也没有报道，这意味着Rp1-D可能是其已知的第一个底物。我们假设，erad介导的活化NLRs降解（EMDAN）是植物中ETI失活的一般机制。我们将使用一系列分子和细胞生物学技术来确定EMDAN和相关途径在控制植物ETI中的重要性，并表征CER9在ERAD中的作用。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

A plant disease resistance response can be triggered by the interaction of nucleotide binding leucine-rich repeat proteins (known as resistance proteins) and specific pathogen-derived proteins known as effectors. This interaction leads to a strong defense response known as effector-triggered immunity (ETI), often resulting in a rapid localized cell death response at the site of pathogen infection. This response must be tightly regulated so it confers resistance, but not excessive so that it inhibits plant growth unnecessarily. Our preliminary evidence suggests that a component of a common protein degradation pathway in maize, ZmCER9, controls degradation of the nucleotide binding leucine-rich repeat proteins, after they are activated. ZmCER9 protein sequence is similar to other proteins involved in a process called endoplasmic reticulum associated degradation (ERAD), which degrades incorrectly-folded proteins. Although ERAD has been characterized in yeast, it is poorly understood in plants. Thus, the interaction of ZmCER9 and subsequent degradation of nucleotide binding leucine-rich repeat proteins appears to be a previously undescribed mechanism that mediates the deactivation of the effector-triggered immunity defense response after activation. We will use a range of techniques to gain insight into how the interaction of CER9 homologs and nucleotide binding leucine-rich repeat proteins control a crucial defense response in plants and identify other proteins involved in this process in plants. This study will contribute to a greater understanding of how plants survive and thrive in the face of biotic stress. This project will also provide training opportunities and STEM enrichment to underserved middle and high school students from low income, rural counties in northeastern, helping to generate the next generation of plant scientists.Plant disease resistance (R-) proteins of the nucleotide binding leucine-rich repeat (NLR) type are activated and induce a strong defense response known as effector-triggered immunity or ETI, upon recognition of specific pathogen-derived effector proteins. ETI’s effectiveness depends on inactivity when the cognate pathogen is absent, rapid induction when a pathogen is recognized and a rapid suppression after induction. The ubiquitin-proteasome pathway, mediated by the sequential actions of E1 (ubiquitin-activating), E2 (ubiquitin-conjugating) and E3 (ubiquitin ligase) enzymes is a major protein modification and degradation pathway found in all eukaryotes. Our preliminary data indicate that maize E3-ligase ZmCER9 mediates degradation of the Rp1-D NLR R-protein, and several other NLR R-proteins, specifically after activation. Based on homology, ZmCER9 is likely a component of the endoplasmic reticulum associated degradation (ERAD), a fundamental eukaryotic quality-control system that degrades incorrectly-folded proteins. ERAD in plants has been poorly characterized and there are no reported substrates of the CER9 complex, meaning that Rp1-D may represent its first known substrate. We hypothesize that ERAD-Mediated Degradation of Activated NLRs (EMDAN) is a general mechanism for the deactivation of ETI in plants. We will use a range of molecular and cell biology techniques to determine the importance of EMDAN and related pathways in controlling ETI in plants and to characterize the role of CER9 in ERAD.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.