Molecular Genetics of Phototransduction

描述（由申请人提供）：我们的研究目的是获得 蛋白激酶C（PKC）如何调节G蛋白偶联 磷脂酶CB介导的信号转导。这种信号机制是 广泛应用于中枢神经系统的作用， 神经递质包括谷氨酸，血清素，乙酰胆碱， 去甲肾上腺素和组胺磷脂酶CB-3的激活导致 产生两种信使：甘油二酯（DAG）和三磷酸肌醇 （1 P3）。IP 3动员细胞内钙，而DAG激活PKC。PKC有 参与信号转导的调节， 细胞过程包括生长、分化和肿瘤发生。 我们将以果蝇的视觉信号转导为模型 系统在视觉级联反应中，E光受体特异性蛋白激酶C 眼蛋白激酶C（eye-PKC）参与了视网膜的脱敏、失活和光适应。 视觉信号。我们之前的研究表明，眼睛蛋白激酶C与一个信号有关， 通过与衔接蛋白INAD相互作用，使转导复合物中的蛋白质发生变化。缺乏 INAD-eyePKC相互作用导致eye-PKC的体内活性丧失。 此外，我们证明了INAD和TRP，也与INAD相关， 被眼PKC磷酸化。 我们将：（1）确定眼PKC磷酸化位点在INAD;（2）研究 INAD的磷酸化如何影响蛋白质-蛋白质相互作用 信号复合物的组成部分;（3）确定眼PKC磷酸化位点 （4）研究TRP的磷酸化如何调节该通道 活性和蛋白质-蛋白质相互作用与INAD;和（5）调查，如果 NORPA也是眼PKC的底物，以及NORPA是如何被眼PKC调节的。 磷酸化 PKC调节细胞功能的许多方面。我们相信我们的研究 PKC对G蛋白偶联信号的调节将为研究 调节神经系统中的细胞间通讯。不受管制 信号传导可能导致神经元的过度活跃或退化，这已经被 与人类的病理生理状况如癫痫有关， 神经退行性疾病

DESCRIPTION (provided by applicant): The objective of our research is to gain insight into how protein kinase C (PKC) regulates G-protein coupled phospholipase CB-mediated signal transduction. This signaling mechanism is widely employed in the central nervous system for the action of several neurotransmitters including glutamate, serotonin, acetylcholine, norepinephrine, and histamine. Activation of phospholipase CB-3 leads to generation of two messengers: diacylglycerol (DAG) and inositol trisphosphate (1P3). IP3 mobilizes intracellular calcium whereas DAG activates PKC. PKC has been implicated in regulation of signal transduction as well as diverse cellular processes including growth, differentiation, and tumorigenesis. We will use the visual transduction in Drosophila melanogaster as a model system. In the visual cascade, E photoreceptor-specific protein kinase C (eye-PKC) is involved in desensitization, deactivation and light adaptation of the visual signaling. We previously showed that eye-PKC is tethered to a signal transduction complex by interacting with an adaptor protein INAD. A lack of the INAD-eyePKC interaction leads to a loss of in vivo activity of eye-PKC. Moreover, we demonstrated that INAD and TRP, which also associate with INAD, are phosphorylated by eye-PKC. We will: (1) identify eye-PKC phosphorylation sites in INAD; (2) investigate how phosphorylation of INAD affects protein-protein interactions with other components of the signaling complex; (3) identify eye-PKC phosphorylation sites in TAP; (4) investigate how phosphorylation of TRP may modulate the channel activities and protein-protein interaction with INAD; and (5) investigate if NORPA is also a substrate of eye-PKC and how NORPA is regulated by phosphorylation. PKC regulates many facets of cellular function. We believe that our research on regulation of G-protein coupled signaling by PKC will provide insight into modulation of cell-cell communications in the nervous system. Un-regulated signaling may lead to hyperactivity or degeneration of neurons, which has been implicated in mans pathophysiological conditions such as epilepsy and neurodegenerative diseases.