Structure-Function Analysis of Type II Metacaspases to Reveal Distinct Activation Mechanisms
II 型元半胱天冬酶的结构功能分析揭示独特的激活机制
基本信息
- 批准号:2052997
- 负责人:
- 金额:$ 135.81万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Continuing Grant
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-09-01 至 2024-08-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Plant stresses caused by diseases, insect pests, heat, cold or drought are major causes of crop loss globally. Understanding how plants respond and recover from stress can greatly facilitate the effort to improve crop productivity through improved breeding and bioengineering approaches. Highly conserved proteases, enzymes that can cleave other proteins, are important in mediating stress resistance as well as developmental pathways in plants. This research will address how plant metacaspases, one type of these conserved proteases, contribute and control the developmental processes associated with stress response. Using advances in visualization techniques of crystallography and cryo-Electron Microscopy techniques, the structures for key members of the plant metacaspase family will be clarified to atomic resolution. In parallel, studies of variants for these metacaspases in transgenic plants will inform on their effects on stress response pathways. Together, these advances will likely have important impacts on agriculture through engineering crops in the future to enhance pathogen resistance and stress tolerance by modifying metacaspase functions. This project will provide diversified, interdisciplinary training to postdoctoral researchers and graduate students to enrich the scientific workforce. In addition, involvement of undergraduate students in experiential training in our multi-disciplinary project should contribute to science education at large. Genetic studies have shown that the metacaspase (MC) proteases are involved in abiotic and biotic stresses-induced cell death in higher plants. Two classes of Type II MCs are exemplified by MC4 and MC9 from Arabidopsis with their activity dependent on calcium ion and mildly acidic pH, respectively. The crystal structure of AtMC4 has been reported at 2.8 Å resolution and uncovered evidence for a multi-step autolytic process in the activation and maturation of the AtMC4 zymogen upon calcium induction. In this project, the structure for AtMC9 will be solved by using crystallography and cryo-Electron Microscopy (cryo-EM), which will enable discovery of the molecular determinants that underlie its activation at acidic pH and independence from calcium. In addition, the structure will be solved for a chimeric protein created from swapping the linker domain between AtMC4 and AtMC9 that resulted in an active proteases which no longer show autolytic cleavage upon its activation by calcium. These new protein structures should help rationalize their observed biochemical differences. To complement these structural studies, in vivo function of mutated and chimeric Type II MCs that have novel combinations of regulatory characteristics will be examined using atmc4 and atmc9 knockout mutants as genetic backgrounds. Lastly, proposed active translocation of AtMC9 zymogen to the acidic apoplastic space upon stresses will be tested by using fusions with acid-stable GFP variants. Together, these in vitro and in vivo trait analysis will reveal the detailed molecular basis for activation of these two metacaspases to induce plant stress response.This award is co-funded by the Cellular Dynamics and Function cluster in the Division of Molecular and Cellular Biosciences and the Plant, Fungal and Microbial Developmental Mechanisms Program in the Division of Integrative Organismal Systems.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.
由病虫害、高温、低温或干旱引起的植物胁迫是全球作物损失的主要原因。 了解植物如何从压力中反应和恢复,可以大大促进通过改进育种和生物工程方法提高作物产量的努力。高度保守的蛋白酶,可以切割其他蛋白质的酶,在介导植物的抗逆性以及发育途径中是重要的。这项研究将解决如何植物metacaspases,这些保守的蛋白酶的一种类型,有助于和控制与应激反应相关的发育过程。利用晶体学和冷冻电子显微镜技术的可视化技术的进步,植物metacaspase家族的关键成员的结构将被澄清到原子分辨率。与此同时,在转基因植物中这些metacaspases的变体的研究将告知其对应激反应途径的影响。总之,这些进展可能会对农业产生重要影响,未来通过改造作物,通过修饰metacaspase功能来增强病原体抗性和胁迫耐受性。该项目将为博士后研究人员和研究生提供多样化的跨学科培训,以丰富科学劳动力。 此外,本科生参与我们的多学科项目的经验培训应有助于科学教育。遗传学研究表明,后半胱天冬酶(MC)蛋白酶参与高等植物非生物和生物胁迫诱导的细胞死亡。两类II型MC的例子是来自拟南芥的MC 4和MC 9,它们的活性分别依赖于钙离子和弱酸性pH。AtMC 4的晶体结构已被报道为2.8 μ m分辨率,并发现了在钙诱导下AtMC 4酶原的激活和成熟中的多步自溶过程的证据。在该项目中,AtMC 9的结构将通过使用晶体学和冷冻电子显微镜(cryo-EM)来解决,这将有助于发现其在酸性pH值下激活并不依赖于钙的分子决定因素。此外,将解析通过交换AtMC 4和AtMC 9之间的接头结构域产生的嵌合蛋白的结构,这导致活性蛋白酶在其被钙激活时不再显示自溶切割。这些新的蛋白质结构应该有助于合理化它们观察到的生化差异。为了补充这些结构研究,将使用atmc 4和atmc 9敲除突变体作为遗传背景,检查具有新的调控特征组合的突变和嵌合II型MC的体内功能。最后,提出的主动易位AtMC 9酶原的酸性质外体空间后,压力将测试通过使用与酸稳定的GFP变体的融合。总之,这些体外和体内性状分析将揭示激活这两种metacaspase诱导植物胁迫反应的详细分子基础。该奖项由分子和细胞生物科学部的细胞动力学和功能集群以及植物,真菌和微生物发育机制计划在综合有机体系统司。这个奖项反映了NSF的法定使命,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。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Eric Lam其他文献
SAFETY AND EFFICACY OF MARGIN AND BASE ABLATION AFTER ENDOSCOPIC MUCOSAL RESECTION OF LARGE NONPEDUNCULATED COLORECTAL POLYPS: A PROSPECTIVE MULTI-CENTER STUDY
内镜下黏膜切除大型无蒂结直肠息肉后切缘及基底消融的安全性和有效性:一项前瞻性多中心研究
- DOI:
10.1016/j.gie.2025.03.840 - 发表时间:
2025-05-01 - 期刊:
- 影响因子:7.500
- 作者:
Edgard Medawar;Mahsa Taghiakbari;Roupen Djinbachian;Neal C. Shahidi;John M. Levenick;Jeffrey D. Mosko;Eric Lam;Christopher Teshima;Matthew Moyer;Simon Bouchard;Érik Deslandres;Heiko Pohl;Daniel Von Renteln - 通讯作者:
Daniel Von Renteln
ENDOSCOPIC ULTRASOUND-GUIDED CHOLEDOCHODUODENOSTOMY: A LEARNING CURVE ANALYSIS BASED ON A MULTICENTRE RANDOMIZED CONTROLLED TRIAL (ELEMENT TRIAL)
- DOI:
10.1016/j.gie.2022.04.1260 - 发表时间:
2022-06-01 - 期刊:
- 影响因子:7.500
- 作者:
Yen-I Chen;Anand Sahai;Gianfranco Donatelli;Eric Lam;Sana Kenshil;Fergal Donnellan;Avijit Chatterjee;Nauzer Forbes;Jeffrey D. Mosko;Etienne Desilets;Sarto C. Paquin;Jennifer J. Telford;Gary R. May;Gurpal S. Sandha;Rachid Mohamed;S. Ian Gan;Andre Roy;Jad Abou Khalil;Corey S. Miller;Jeffrey Barkun;Alan Barkun - 通讯作者:
Alan Barkun
Structural determinants for pH-dependent activation of a plant metacaspase
植物类胱天蛋白酶 pH 依赖性激活的结构决定因素
- DOI:
10.1038/s41467-025-60253-y - 发表时间:
2025-05-29 - 期刊:
- 影响因子:15.700
- 作者:
Haijiao Liu;Max Henderson;Zhili Pang;Qingfang Zhang;Eric Lam;Qun Liu - 通讯作者:
Qun Liu
Programmed cell death, mitochondria and the plant hypersensitive response
程序性细胞死亡、线粒体与植物过敏反应
- DOI:
10.1038/35081184 - 发表时间:
2001-06-14 - 期刊:
- 影响因子:48.500
- 作者:
Eric Lam;Naohiro Kato;Michael Lawton - 通讯作者:
Michael Lawton
REAL WORLD EFFICACY OF A COLONIC SELECTIVE RESECTION ALGORITHM FOR LARGE NON-PEDUNCULATED COLON POLYPS
- DOI:
10.1016/j.gie.2024.04.2709 - 发表时间:
2024-06-01 - 期刊:
- 影响因子:
- 作者:
Shirley Jiang;Aein Zarrin;Angad Walia;Cherry Galorport;Robert Enns;Eric Lam;Neal Shahidi - 通讯作者:
Neal Shahidi
Eric Lam的其他文献
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{{ truncateString('Eric Lam', 18)}}的其他基金
Targeting the FOXM1 signature for early diagnosis and treatment in cholangiocarcinoma
靶向 FOXM1 特征用于胆管癌的早期诊断和治疗
- 批准号:
MR/N012097/1 - 财政年份:2016
- 资助金额:
$ 135.81万 - 项目类别:
Research Grant
Function and Regulation of Metacaspases in Plant Cell Death
后半胱天冬酶在植物细胞死亡中的功能和调节
- 批准号:
1258071 - 财政年份:2013
- 资助金额:
$ 135.81万 - 项目类别:
Continuing Grant
Meeting: International Conference on Duckweed Research at Rutgers University on August 21-24, 2013
会议:2013 年 8 月 21-24 日在罗格斯大学举行的浮萍研究国际会议
- 批准号:
1338642 - 财政年份:2013
- 资助金额:
$ 135.81万 - 项目类别:
Standard Grant
IGERT: Solutions for Renewable and Sustainable Fuels in the 21st Century
IGERT:21 世纪可再生和可持续燃料解决方案
- 批准号:
0903675 - 财政年份:2009
- 资助金额:
$ 135.81万 - 项目类别:
Continuing Grant
Functional Analysis of Metacaspases in Plant Development and Cell Death.
植物发育和细胞死亡中元半胱天冬酶的功能分析。
- 批准号:
0744709 - 财政年份:2008
- 资助金额:
$ 135.81万 - 项目类别:
Continuing Grant
Chromatin Charting: Organization and Dynamics of Plant Nuclear DNA in situ
染色质图谱:植物核 DNA 的原位组织和动态
- 批准号:
0077617 - 财政年份:2000
- 资助金额:
$ 135.81万 - 项目类别:
Continuing Grant
Chromatin Charting: Organization and Dynamics of Plant Nuclear DNA in situ
染色质图谱:植物核 DNA 的原位组织和动态
- 批准号:
9872636 - 财政年份:1998
- 资助金额:
$ 135.81万 - 项目类别:
Standard Grant
NSF Postdoctoral Research Fellowships in Plant Biology
NSF 植物生物学博士后研究奖学金
- 批准号:
8508764 - 财政年份:1985
- 资助金额:
$ 135.81万 - 项目类别:
Fellowship Award
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