Interaction of ricin A chain with the ribosomal stalk

蓖麻毒素 A 链与核糖体柄的相互作用

• 批准号: 7942717
• 负责人: NILGUN E TUMER
• 金额: $ 6.27万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7942717
• 关键词: Apoptosis; Archaea; Architecture; Bacteria; Binding; Biochemical; Biological Models; Cell Death; Cells; Collaborations; Complex; Data; Defect; Depurination; Development; Docking; Environment; Glycine decarboxylase; Grant; Human; Induction of Apoptosis; Link; MAPK14 gene; MAPK8 gene; Mammalian Cell; Messenger RNA; Methods; Modeling; Molecular; Molecular Probes; Mutagenesis; Myelin P2 Protein; Nature; New Brunswick; New Jersey; Parents; Poland; Preparation; Procedures; Protein Biosynthesis; Proteins; RNA Splicing; Research; Resistance; Resources; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Ricin; Ricin A Chain; Role; Saccharomyces; Shiga-Like Toxins; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Spain; Specificity; Structure; System; Time; United States National Institutes of Health; Universities; Yeasts; base; cytotoxicity; genetic analysis; in vivo; insight; mutant; novel; parent grant; pokeweed antiviral protein; prevent; tool

DESCRIPTION (provided by applicant): This is a FIRCA application for the collaboration between Dr. Nilgun Tumer at Rutgers University, New Brunswick, New Jersey and Dr. Marek Tchsrzewski at the Maria Curie Sklodowska University in Poland. This research will be done at the Maria Curie Sklodowska University in Poland as an extension of NIH Grant No. R01-AI072425, 3-15-2007 to 2-29-2012. This collaboration makes use of the unique resources and research environment in Dr. Tchsrzewski's lab to use ricin A chain (RTA) as a molecular tool to probe the function of the eukaryotic ribosomal stalk and to identify the functional part of the stalk structure. In spite of its fundamental importance in protein synthesis, the molecular architecture of the eukaryotic stalk and details of its function are not known due to the lack of appropriate molecular tools. Dr. Tchsrzewski has developed the yeast strains and the methods to isolate the in vivo assembled eukaryotic ribosomal stalk for the first time. Isolation of the whole native stalk from yeast has revolutionized the method of stalk preparation and facilitated research on the function of the eukaryotic ribosomal stalk. Dr. Tumer has developed the model systems to study the activity of ricin A chain (RTA) and discovered that RTA binds to the ribosomal stalk to inhibit protein synthesis. In collaboration with Dr. Tchsrzewski, Dr. Tumer showed direct interaction between RTA and the isolated native yeast ribosomal stalk, and obtained preliminary data demonstrating that RTA can be used as a very specific molecular tool to probe the structure and the function of the eukaryotic stalk. We propose to identify the stalk components critical for binding RTA, study the interaction of RTA with stalk complexes from different species, including bacteria, archaea, yeast and human and to identify the functional part of the stalk structure. This collaboration will connect the ongoing structure function analysis of RTA in the parent grant with biochemical, structural and genetic analysis of the stalk in Dr. Tchsrzewski's lab in Poland and will provide new insights into the role of the ribosomal stalk in protein synthesis. PUBLIC HEALTH RELEVANCE: This project seeks to probe the function of the eukaryotic ribosomal stalk and to identify the functional part of the stalk structure using ricin A chain as a molecular probe. It will provide a better understading of the stalk structure and its role in protein synthesis.

描述（由申请人提供）：这是新泽西州新不伦瑞克市罗格斯大学的Nilgun Tumer博士和波兰玛丽亚居里大学的Marek Tchsrzewski博士合作的FIRCA申请。这项研究将在波兰的玛丽亚·居里·斯克洛多夫斯卡大学进行，作为美国国立卫生研究院拨款号的延伸。R01-AI072425, 3-15-2007至2-29-2012。此次合作利用Tchsrzewski博士实验室独特的资源和研究环境，利用蓖麻毒素A链（ricin A chain， RTA）作为分子工具，探测真核核糖体茎秆的功能，并鉴定茎秆结构的功能部分。尽管它在蛋白质合成中具有重要的基础作用，但由于缺乏适当的分子工具，真核生物柄的分子结构及其功能细节尚不清楚。Tchsrzewski博士首次开发了酵母菌株和分离体内组装真核核糖体茎的方法。从酵母中分离出完整的天然酵母柄，彻底改变了酵母柄的制备方法，促进了对真核核糖体秸秆功能的研究。Tumer博士开发了模型系统来研究蓖麻毒素A链（RTA）的活性，并发现RTA与核糖体柄结合以抑制蛋白质合成。在与Tchsrzewski博士的合作中，Tumer博士展示了RTA与分离的天然酵母核糖体茎秆之间的直接相互作用，并获得了初步数据，证明RTA可以作为一种非常特殊的分子工具来探测真核生物茎秆的结构和功能。我们建议鉴定与RTA结合至关重要的茎秆组分，研究RTA与不同物种（包括细菌、古生菌、酵母和人类）茎秆复合物的相互作用，并鉴定茎秆结构的功能部分。此次合作将把亲本基金中正在进行的RTA结构功能分析与Tchsrzewski博士在波兰的实验室中对RTA进行的生化、结构和遗传分析联系起来，并将为核糖体茎秆在蛋白质合成中的作用提供新的见解。