Phospholipase D in phototransduction and cellularization

描述（由申请人提供）：我们研究的长期目标是了解磷脂酶D（PLD）的分子机制：1）调节对光转导至关重要的G蛋白偶联信号级联反应，2）介导高尔基体衍生囊泡的重定向，用于果蝇细胞化过程中的膜生物合成。PLD水解磷脂酰胆碱产生脂质信使磷脂酸（PA）。在哺乳动物系统中，PLD具有多种功能，包括囊泡运输、膜形成、细胞骨架重排和胞吐作用。在酵母中，PLD在减数分裂前孢子膜形成过程中的膜生物发生中起着关键作用。具体目的如下：1）确定dPLD在眼内的亚细胞定位，并通过功能获得和功能丧失分析来探索其作用; 2）利用突变互补从遗传学角度剖析dPLD在光转导中的功能; 3）利用标记共定位、突变互补和功能丧失分析来确定PLD在细胞化中的功能。

DESCRIPTION (provided by applicant): The long-term goal of our research is to understand the molecular mechanisms by which phospholipase D (PLD) 1) regulates the G-protein coupled signaling cascade critical for phototransduction and 2) mediates the redirection of Golgi-derived vesicles for membrane biogenesis during cellularization in Drosophila melanogaster. PLD hydrolyzes phosphatidylcholine to generate the lipid messenger, phosphatidic acid (PA). In mammalian systems, PLD has diverse functions, including vesicle trafficking, membrane formation, cytoskeletal rearrangements, and exocytosis. In yeast, PLD plays a critical role in membrane biogenesis during meiotic prospore membrane formation. The Specific Aims of this proposal are as follows: 1) determine the sub-cellular localization of dPLD in the eye and probe its role through gain- and loss-of-function analysis, 2) use mutant complementation to dissect genetically the function of dPLD in phototransduction, and 3) use marker co-localization, mutant complementation, and loss-of-function analysis to determine the function of PLD in cellularization.