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（由申请人提供）：激发-转录（E-T）耦合是将细胞的电或化学激活转化为传递到细胞核的信号的过程，通过这种方式，可以以活性调节基因的表达。所产生的神经元重塑被认为对于神经发育、学习和记忆以及药物成瘾过程中的长期适应性变化是必要且重要的。 E-T 耦合最受关注的例子是 Ca2+ 信号传导。转录因子 CREB（cAMP 反应元件结合）蛋白通过 Ser133 磷酸化作为 Ca2+ 流入的重要来源，电压门控 Ca2+ 通道的生物物理和生化特性已得到充分研究，在 E-T 耦合中似乎 Ca2+ 流入。通过不同的 Ca2+ 通道可以参与不同的信号通路到达细胞核，例如，CaV1（也称为 L 型）通道比 CaV2 通道具有很大的优势，尽管 CaV1 通道。我们最近的《细胞》论文发现，这种效力差异可以通过两类 Ca2+ 通道如何使用局部和全局 Ca2+ 信号传导的差异来解释。 CaV1 通道的纳米域优势尚不清楚。现在我们准备提供关键问题的详细表征：什么将长距离信号从 CaV1 锚定的信号复合物传递到细胞核？答案：Ca2+/CaM 向细胞核的易位取决于我们现在识别为 γCaMKII 的协同转运蛋白。该穿梭机收集细胞质 Ca2+/CaM，在核定位信号的控制下将其隔离在 CaV1 通道上。信号传导机制依赖于 γCaMKII、βCaMKII 和 CaN，这些信号分子在 CaV1 纳米结构域中起作用，并且与多种神经精神疾病有关该提案的重点是了解 γCaMKII/CaM 易位的细胞机制，并提出了三个具体目标（1）定义将 CaV1 活性与核 CREB ​​磷酸化和 CRE 依赖性转录联系起来的 Ca2+ 信号传导机制。实时易位并评估通过这种穿梭机制传递到细胞核的 Ca2+/CaM 的影响 (2) 我们将使用遗传构建体操纵 γCaMKII 途径。确定 CREB ​​磷酸化所需的分子成分，我们将改变该通路关键步骤中涉及 CaM、βCaMKII、CaN 和 PP2A 的结合相互作用和酶作用。 (3) 了解新皮质神经元中依赖 CaV1 的 CaM 穿梭并定义不同的作用。纳米结构域 Ca2+ 信号传导和电压门控构象信号传导的 E-T 耦合的研究 更清楚地了解 CaV1 通道和 CREB ​​信号传导之间的联系将产生有利的影响。理解基因表达的变化如何改变神经网络中神经元的功能因此，这项研究与基本细胞相关。 生物学和各种疾病状态，如成瘾、自闭症和其他神经精神疾病。