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

9603360弗罗曼该奖项支持纽约州立大学石溪分校的迈克尔·弗罗曼教授和日本岐阜市大学医学院的野泽良典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，磷脂酶D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)利用定点突变继续结构-功能实验，以确定与辅因子如蛋白激酶C和小G蛋白相互作用所需的氨基酸，以及细胞内的定位；以及2)确定PLD1或2是否通过刺激特征良好的细胞系中的特定信号转导途径而被激活，以及这种激活是否是必要的或足以产生后续的细胞反应，通常被归因于PLD的激活。该项目汇集了两个实验室的努力，这两个实验室构成了这一进程的不同部分。美国人在标准和先进的分子生物学技术、向细胞内显微注射质粒和蛋白质、胚胎原位杂交和转基因小鼠技术方面拥有丰富的经验。十年来，日本人一直在使用各种方法来研究PLD，并在各种药理PLD激活剂和抑制剂以及测量与PLD激活相关的细胞后果，如5-羟色胺分泌、肥大细胞脱颗粒和二酰甘油产生方面拥有丰富的经验。在许多不同种类的细胞和组织中，PLD的激活与各种正常的生理和疾病过程有关。研究结果将为不同模型体系的优势和局限性提供现实的展望。这将作为S决定应该实施哪些模式的基础。通过思想和技术的交流，这个项目将扩大我们的基础知识基础，促进国际理解与合作。***

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. ***