Interaction of ricin A chain with the ribosomal stalk

项目摘要 这是一个FIRCA的应用程序， 新泽西州新玩法大学和马雷克·切泽夫斯基博士在玛丽亚·居里 波兰的斯克洛多夫斯卡大学。这项研究将在玛丽亚·居里·斯克洛多夫斯卡 波兰大学，作为NIH批准号R 01-AI 072425的扩展，2007年3月15日至2012年2月29日。 这项合作利用了博士独特的资源和研究环境。 Tch <$rzewski的实验室使用蓖麻毒素A链（RTA）作为分子工具来探测 真核生物核糖体柄，并确定柄结构的功能部分。尽管其 蛋白质合成的基础重要性，真核生物茎的分子结构 由于缺乏合适的分子工具，其功能的细节尚不清楚。博士 Tch <$rzewski已经开发了酵母菌株和分离体内组装的方法， 真核生物的核糖体柄。从酵母中分离整个天然茎， 彻底改变了秸秆制备方法，促进了对秸秆功能的研究， 真核核糖体柄。Tumer博士开发了模型系统来研究 蓖麻毒素A链（RTA），并发现RTA结合到核糖体柄，以抑制蛋白 合成.在与Tch <$rzewski博士的合作中，Tumer博士展示了 RTA和分离的天然酵母核糖体茎，并获得了初步数据，证明 RTA可以作为一种非常特异的分子工具来探测 真核生物的茎我们建议确定对RTA结合至关重要的茎成分，研究 RTA与来自不同物种的茎复合物的相互作用，包括细菌，古细菌， 酵母和人类，并确定茎结构的功能部分。这项合作将 将正在进行的RTA结构功能分析与生物化学结合起来， 在波兰Tch ́ rzewski博士的实验室里对茎的结构和遗传分析， 对核糖体柄在蛋白质合成中作用的新认识。

PROJECT SUMMARY This is a FIRCA application for the collaboration between Dr. Nilgun Tumer at Rutgers University, New Brunswick, New Jersey and Dr. Marek Tch¿rzewski 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. Tch¿rzewski'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. Tch¿rzewski 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. Tch¿rzewski, 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. Tch¿rzewski's lab in Poland and will provide new insights into the role of the ribosomal stalk in protein synthesis.