U.S.-Japan Cooperative Science: Regulation and Roles of Signal Transducing

9603360 Frohman该奖项支持纽约州立大学石溪分校（SUNY）的Michael Frohman教授和日本岐阜大学医学院的yoshiori Nozawa教授之间为期两年的合作研究项目。该项目将研究磷脂酶D （PLD）的生化、细胞和生理作用，这种酶被认为参与信号转导、囊泡运输和细胞分裂的控制。该项目的主要目标是：1)继续使用定点诱变进行结构-功能实验，以确定与辅助因子（如蛋白激酶C和小G-蛋白）相互作用所需的氨基酸，以及细胞内定位；2)确定在特征明确的细胞系中，PLD1或2是否通过刺激特定的信号转导途径而被激活，以及这种激活对于产生通常归因于PLD激活的后续细胞反应是否必要或充分。该项目汇集了构成该过程不同部分的两个实验室的努力。美国人在标准和先进的分子生物学技术、向细胞中微量注射质粒和蛋白质、胚胎原位杂交和转基因小鼠技术方面拥有丰富的经验。日本人研究PLD的方法已有十年之久，在多种PLD药理激活剂和抑制剂以及测量与PLD激活相关的细胞后果（如血清素分泌、肥大细胞脱颗粒和二酰基甘油生成）方面拥有丰富的经验。PLD的激活与许多不同种类的细胞和组织中的各种正常生理和疾病过程有关。研究结果将对不同模型系统的优势和局限性提供现实的期望。这将作为确定应该实现哪些模型的基础。通过思想和技术的交流，该项目将扩大我们的基础知识基础，促进国际了解与合作。＊＊＊

9603360 Frohman This award supports a two year collaborative research project between Professor Michael Frohman of the State University of New York (SUNY) at Stony Brook and Professor Yoshinori Nozawa of the Gifu University School of Medicine in Japan. The project will investigate the biochemical, cellular, and physiological roles of Phospholipase D (PLD), an enzyme thought to be involved in signal transduction, vesicular trafficking, and the control of cell division. The major goals of this project are: 1) to continue structure-function experiments using site-directed mutagenesis to identify amino acids required for interaction with cofactors such as protein kinase C and small G- proteins, and intracellular localization; and 2) to determine whether PLD1 or 2 are activated by stimulation of specific signal transduction pathways in well characterized cell lines, and whether such activation is necessary or sufficient to generate the subsequent cellular responses that have been ascribed to PLD activation in general. The project brings together the efforts of two laboratories that have constituted different parts of the process. The Americans have extensive experience with standard and advanced molecular biology techniques, microinjection of plasmids and proteins into cells, embryonic in situ hybridization, and transgenic mouse technology. The Japanese have been using approaches to study PLD for a decade and have extensive experience with a wide variety of pharmacological PLD activators and inhibitors, and with measuring the cellular consequences correlated with PLD activation, such as serotonin secretion, mast cell degranulation, and diacylglycerol production. PLD activation has been linked to a wide variety of normal physiological and disease processes in many different kinds of cells and tissues. Results of the research will provide realistic expectation about the advantages and limitation of the different model systems. This would serve as the basi s to determine which models should be implemented. Through the exchange of ideas and technology, this project will broaden our base of basic knowledge and promote international understanding and cooperation. ***