Molecular Genetics of Phototransduction

光转导的分子遗传学

• 批准号: 6624209
• 负责人: BIH-HWA SHIEH
• 金额: $ 33.98万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6624209
• 关键词: Drosophilidae; G protein; biological signal transduction; calcium channel; central nervous system; diacylglycerols; enzyme activity; genetically modified animals; inositol phosphates; intermolecular interaction; mass spectrometry; matrix assisted laser desorption ionization; molecular genetics; phosphorylation; photoactivation; protein kinase C; protein protein interaction; protein structure function; receptor coupling; recombinant proteins; site directed mutagenesis; visual photoreceptor; visual phototransduction

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.

描述(申请人提供)：我们研究的目的是获得 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 在中枢神经系统中被广泛用于几种 神经递质包括谷氨酸、5-羟色胺、乙酰胆碱、 去甲肾上腺素和组胺。磷脂酶CB-3的激活导致 两种信使的产生：二酰甘油(DAG)和三磷酸肌醇 (1P3)。IP3动员细胞内钙离子，而DAG激活PKC。PKC有 与信号转导的调节有关，也与多种 包括生长、分化和肿瘤发生在内的细胞过程。 我们将以果蝇的视觉传导为模型 系统。在视觉级联中，E光感受器特异的蛋白激酶C (Eye-PKC)参与大脑皮层的脱敏、失活和光适应。 视觉信号。我们之前已经证明了Eyes-PKC与一个信号有关 通过与接头蛋白inad相互作用而形成的转导复合体。缺乏可持续的 内-眼PKC相互作用导致眼-PKC在体内的活性丧失。 此外，我们还证明了同样与INAD相关的INAD和Trp， 被Eye-PKC磷酸化。 我们将：(1)确定INAD中眼睛-PKC的磷酸化位点；(2)研究 Inad的磷酸化如何影响蛋白质与其他蛋白质的相互作用 信号复合体的组成；(3)确定Eyes-PKC磷酸化位点 在TAP中；(4)研究Trp的磷酸化如何调节通道 与INAD的活性和蛋白质-蛋白质相互作用；以及(5)研究 Norpa也是Eye-PKC的底物，以及Norpa是如何被 磷酸化。 PKC调节细胞功能的许多方面。我们相信我们对此的研究 PKC对G蛋白偶联信号的调节将为深入了解 神经系统中细胞间通讯的调制。不受监管 信号可能会导致神经元过度活跃或变性，这已经被 与MAN的病理生理状况有关，如癫痫和 神经退行性疾病。