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）：这是一个FIRCA应用程序之间的合作博士Nilgun Tumer在罗格斯大学，新玩法，新泽西和博士Marek Tchsrzewski在玛丽亚居里Sklodowska大学在波兰。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NIH批准号R 01-AI 072425（2007年3月15日至2012年2月29日）的扩展。这项合作利用Tchsrzewski博士实验室的独特资源和研究环境，使用蓖麻毒素A链（RTA）作为分子工具来探测真核生物核糖体茎的功能，并识别茎结构的功能部分。尽管它在蛋白质合成中具有根本的重要性，但由于缺乏适当的分子工具，真核茎的分子结构及其功能的细节尚不清楚。Tchsrzewski博士首次开发了酵母菌株和分离体内组装的真核核糖体柄的方法。从酵母中分离出完整的天然柄，彻底改变了柄的制备方法，并促进了真核生物核糖体柄功能的研究。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之间的直接相互作用，并获得了初步数据，证明RTA可用作一种非常特异的分子工具来探测真核生物茎的结构和功能。我们建议确定的茎组件结合RTA的关键，研究RTA与不同物种，包括细菌，古细菌，酵母和人类的茎复合物的相互作用，并确定茎结构的功能部分。这项合作将把正在进行的RTA结构功能分析与波兰Tchsrzewski博士实验室的茎生物化学、结构和遗传分析联系起来，并将为核糖体茎在蛋白质合成中的作用提供新的见解。 公共卫生相关性：本研究以蓖麻毒素A链为分子探针，探讨真核生物核糖体柄的功能，并鉴定柄结构的功能部分。这将有助于更好地理解茎结构及其在蛋白质合成中的作用。