Calcium Channels, Calmodulin and Nuclear CREB Signaling

钙通道、钙调蛋白和核 CREB ​​信号传导

• 批准号: 9306042
• 负责人: RICHARD W TSIEN
• 金额: $ 40.26万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9306042
• 关键词: Action Potentials; Address; Alpha Cell; Autistic Disorder; Binding; Biochemical; Biological Neural Networks; Biophysics; Brain; Calcium Channel; Calmodulin; Cell Nucleus; Cells; Cellular biology; Chelating Agents; Chemicals; Chimeric Proteins; Communication; Complex; Couples; Coupling; Cyclic AMP Response Element; Cyclic AMP-Responsive DNA-Binding Protein; Cytoplasm; DNA Sequence Alteration; Development; Diffusion; Disabled Persons; Disease; Drug Addiction; Egtazic Acid; Enzymes; Event; Family; Frequencies; Funding; Gene Expression; Genetic; Genetic Transcription; Goals; Health; Image; Income; Label; Learning; Ligands; Link; Membrane; Memory; Minority; Modification; Molecular; Molecular Conformation; Movement; Nail plate; Neurons; Nuclear; Nuclear Localization Signal; Oral cavity; Paper; Pathway interactions; Peripheral; Phosphorylation; Phosphotransferases; Play; Privatization; Process; Property; Protein Dephosphorylation; Protein Phosphatase 2A Regulatory Subunit PR53; Radial; Research; Resistance; Role; Scheme; Secure; Serine; Signal Pathway; Signal Transduction; Signaling Molecule; Source; Stimulus; Sum; Surface; System; Testing; Time; Tissues; Travel; Vision; Work; addiction; base; calmodulin-dependent protein kinase II; cell type; excitatory neuron; knock-down; member; mutant; neocortical; neuron development; neuropsychiatric disorder; novel; operation; protein expression; public health relevance; response; small hairpin RNA; symporter; transcription factor; voltage

 DESCRIPTION (provided by applicant): Excitation-transcription (E-T) coupling is a process that converts the electrical or chemical activation of a cell to a signal conveyed to the nucleus. n this way, the expression of genes can be modulated in an activity-dependent manner. The neuronal remodeling that results is recognized to be necessary and important for long-term adaptive changes during neuronal development, learning and memory and drug addiction. The most scrutinized example of E-T coupling is Ca2+ signaling to the transcription factor CREB (cAMP response element-binding) protein via phosphorylation at Ser133. As an important source of Ca2+ influx, voltage-gated Ca2+ channels have been well studied for their biophysical and biochemical properties. Interestingly, in E-T coupling it seems that Ca2+ influxes through different Ca2+ channels can engage different signaling pathways to the nucleus. For example, CaV1 (also called L-type) channels enjoy a big advantage over CaV2 channels, even though CaV1 channels contribute only a minority of the overall Ca2+ entry in neurons. Our recent Cell paper uncovered that this disparity in potency can be explained by differences in how the two classes of Ca2+ channels employ local and global Ca2+ signaling. However, the 'private line' for the nanodomain advantage of CaV1 channels is unclear. Now we are poised to provide a detailed characterization of the critical question: what carries the long-distance signal from CaV1-anchored signaling complex to the nucleus? We have an answer: Ca2+/CaM translocation to the nucleus depends on a co-transporter that we now identify as γCaMKII. This shuttle gathers cytoplasmic Ca2+/CaM, sequestering it at the CaV1 channel before traveling to the nucleus under control of a nuclear localization signal. This signaling mechanism relies on γCaMKII, βCaMKII and CaN, signaling molecules that operate in the CaV1 nanodomain and also have been implicated in multiple neuropsychiatric diseases. This proposal focuses on understanding the cellular machinery of γCaMKII/CaM translocation and three specific aims are proposed. (1) Define the dynamics of Ca2+ signaling mechanisms that link CaV1 activity to nuclear CREB phosphorylation and CRE-dependent transcription. We will track γCaMKII translocation in real time and assess the impact of Ca2+/CaM delivered to the nucleus via this shuttle mechanism. (2) We will manipulate the γCaMKII pathway using genetic constructs in order to nail down the molecular components required for CREB phosphorylation. We will alter binding interactions and enzymatic actions involving CaM, βCaMKII, CaN, and PP2A at critical steps along the pathway. (3) Understand CaV1-dependent CaM shuttling in neocortical neurons and define distinct roles of nanodomain Ca2+ signaling and voltage gated conformational signaling for E-T coupling. Gaining a clearer picture of the linkage between CaV1 channels and CREB signaling will have a favorable impact on understanding how changes in gene expression alter the function of neurons in neural networks. Thus, the research is relevant both to basic cell biology and to disease states as diverse as addiction, autism and other neuropsychiatric diseases.

 描述（由应用提供）：激发 - 转录（E-T）耦合是一个将细胞的电或化学激活转换为传递到核us的信号的过程。 n这样，可以以活性依赖性方式调节基因的表达。确认结果的神经元重塑对于在神经元发展，学习和记忆和药物成瘾过程中的长期适应性变化是必要和重要的。 E-T耦合的最严格审查的例子是通过Ser133处的磷酸化来向转录因子CREB（CAMP响应元件结合）蛋白进行CA2+信号传导。作为CA2+影响的重要来源，电压门控的Ca2+通道一直是其生物物理和生化特性的很好的研究。有趣的是，在E-T耦合中，Ca2+通过不同的CA2+通道的影响似乎可以使不同的信号通路传入到核。例如，即使CAV1通道仅贡献神经元中总体CA2+进入的少数，CAV1（也称为L型）通道比CAV2通道具有很大的优势。我们最近的细胞纸发现，在两种类别的Ca2+通道员工本地和全局CA2+信号传导方面的差异可以通过差异来解释。但是，CAV1通道的纳米域优势的“私有线”尚不清楚。现在，我们被毒死了一个关键问题的详细表征：什么是带有CAV1锚定信号复合物到核的长距离信号？我们有一个答案：Ca2+/CAM转移到细胞核的取决于我们现在将其识别为γCAMKII的共发器。这种班车收集了细胞质Ca2+/cam，在控制核定位信号的核之前将其隔离在CAV1通道上。这种信号传导机制依赖于γCAMKII，βCAMKII和CAN，即在CAV1纳米域中运行的信号分子，并且也已在多种神经精神疾病中实施。该建议的重点是理解γCAMKII/CAM易位的细胞机制，并提出了三个特定目标。 （1）定义CA2+信号传导机制的动力学，这些动力学将CAV1活性与核CREB磷酸化和CRE依赖性转录联系起来。我们将实时跟踪γCAMKII易位，并评估通过这种班车机制传递给核用户的Ca2+/CAM的影响。 （2）我们将使用遗传构建体操纵γCAMKII途径，以钉住CREB磷酸化所需的分子成分。我们将在沿途径的关键步骤上改变涉及CAM，βCAMKII，CAN和PP2A的结合相互作用和酶促作用。 （3）了解新皮质神经元中的CAV1依赖性凸轮穿梭，并定义纳米域Ca2+信号传导和电压门控构象信号传导的不同作用，以进行E-T偶联。更清楚地了解CAV1通道和CREB信号之间的联系将对理解基因表达的变化如何改变神经元中神经元的功能有利。这是与基本细胞有关的研究 生物学和疾病，如成瘾，自闭症和其他神经精神疾病。