Interaction of ricin A chain with the ribosomal stalk

项目总结 这是FIRCA申请罗格斯大学尼尔贡·图默博士之间的合作 新泽西州新不伦瑞克大学和玛丽亚居里夫人的Marek Tch？Rzewski博士 波兰的斯克罗多夫斯卡大学。这项研究将在玛丽亚·居里·斯克罗多夫斯卡进行 波兰大学，作为国家卫生研究院第号补助金的延伸。R01-AI072425,2007年3月15日至2012年2月29日。 这一合作利用了DR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)作为分子工具来探索 并鉴定真核核糖体秸秆结构的功能部分。尽管它有 真核生物茎的分子结构在蛋白质合成中的重要作用 由于缺乏合适的分子工具，其功能的细节尚不清楚。Dr。 Tch？Rzewski开发了酵母菌株和分离体内组装的 首次发现真核核糖体柄。从酵母菌中分离出整个本地秸秆 革新了秸秆制备方法，促进了对玉米秸秆功能的研究 真核核糖体的茎。Tumer博士已经开发了模型系统来研究 蓖麻毒素A链(RTA)，并发现RTA与核糖体茎结合以抑制蛋白质 综合。在与TchéRzewski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其结构和功能 真核生物的茎。我们建议确定对结合RTA至关重要的茎成分，研究 RTA与不同物种的茎复合体的相互作用，包括细菌，古细菌， 酵母菌和人类，并鉴定茎结构的功能部分。此次合作将 将母基金中正在进行的RTA结构功能分析与生化相结合， 对Tch？Rzewski博士在波兰的实验室中的茎的结构和遗传分析，并将提供 对核糖体茎在蛋白质合成中的作用的新见解。

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.