Molecular Mechanisms of Signal Transduction Involving Light, Redox and Transmembrane Complexes

涉及光、氧化还原和跨膜复合物的信号转导的分子机制

• 批准号: 9276852
• 负责人: BRIAN R CRANE
• 金额: $ 44.07万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9276852
• 关键词: Amaze; Animal Model; Architecture; Behavior; Behavioral Assay; Biological; Borrelia; Cells; Chemicals; Chemoreceptors; Chemotaxis; Cholera; Circadian Rhythms; Communication; Complex; Coupling; Cryoelectron Microscopy; Defect; Devices; Diabetes Mellitus; Disease; Drosophila melanogaster; Electron Spin Resonance Spectroscopy; Environment; Feedback; Flagella; Functional disorder; Fungal Proteins; Gene Expression; Generations; Genes; Genetic; Genetic Transcription; Goals; Helicobacter pylori; Human; Immune system; Infection; Invaded; Investigation; Ions; Light; Lyme Disease; Macromolecular Complexes; Malignant Neoplasms; Manic; Memory; Mental Depression; Mental disorders; Metabolic; Metabolic Diseases; Metabolism; Metals; Molecular; Molecular Conformation; Motion; Motor; Nature; Neurospora crassa; Obesity; Optics; Organism; Output; Oxidation-Reduction; Phase; Physiologic pulse; Prokaryotic Cells; Proteins; Regulation; Repressor Proteins; Roentgen Rays; Schizophrenia; Sensory; Signal Transduction; Sleep Disorders; Spectrum Analysis; Structure; Switching Complex; Syphilis; System; Techniques; Tissues; Torque; Transcription Coactivator; Treponema pallidum; Ulcer; Vibrio cholerae; X-Ray Crystallography; base; biophysical analysis; biophysical techniques; cell growth; cell motility; circadian pacemaker; flavin nucleotide; fly; infectious disease treatment; insight; malignant stomach neoplasm; nanomachine; pathogen; programs; protein-histidine kinase; receptor; response; treatment strategy

The Crane group studies mechanisms of signal transduction with the overall goal of understanding behavior at the molecular level. This understanding will be achieved by defining the structure and dynamics of key macromolecular complexes that coordinate gene expression and transmembrane signaling in two systems that rely on highly cooperative interactions to respond to light, redox and chemical environment. The first, bacterial chemotaxis, concerns the motion of prokaryotic cells toward external stimulants. Chemotaxis is a paradigm for understanding transmembrane communication, intracellular information transfer, and motility. Importantly, many human pathogens that cause diseases such as cholera, gastric cancer and lyme rely on chemotaxis to establish infection. The sensory apparatus underlying chemotaxis, hereafter called “the chemosome”, displays amazing sensitivity, dynamic range and a rudimentary molecular memory. In the chemosome, receptors, histidine kinases (CheA) and coupling proteins assemble into a specific architecture, whose details are just emerging. This proposal continues efforts to understand chemosome assembly, chemoreceptor conformational signaling, and ultimately, CheA regulation through restructuring of the receptor arrays. Chemosome output modulates Nature's consummate nanomachine – the flagella motor. The ultrastructure of the switch complex within the motor will be defined to understand torque generation, direction switching and response to chemosome signals. The second system, eukaryotic circadian clocks, comprises cell-autonomous timing devices that pace metabolism to the diurnal cycle. Clocks are composed of transcriptional-translational feedback loops (TTFLs) within which repressor proteins inhibit the transcriptional activators of their own genes. Light entrains the clock phase by stimulating photosensors that impinge directly on the TTFLs. In humans, aberrant clock function causes mental illness (sleep disorders, depression, mania), cell growth deregulation (cancer) and metabolic defects (diabetes and obesity). This project proposes structural and mechanistic investigations of the key repressor and light-setting activities common to clocks in higher organisms. Biophysical studies will be conducted on the circadian proteins of fungi (Neurospora crassa) and flies (Drosophila melanogaster). Both model organisms provide genetic systems and behavioral assays to probe the biological relevance of mechanistic insights. A complimentary set of techniques including X-ray crystallography, small-angle X-ray scattering, optical spectroscopy, cryo-electron microscopy and pulse-dipolar ESR spectroscopy (PDS) will be applied to accomplish these goals. For PDS, new strategies for incorporating spin probes based on nitroxides, flavins, nucleotides, and metal ions will be developed and deployed. Overall, this program aims to provide a molecular understanding for sensing and response in bacterial chemotaxis and eukaryotic circadian rhythms through the synergistic application of biophysical methods.

起重机小组研究信号转导机制，其总体目标是了解行为， 分子水平。这种理解将通过定义键的结构和动力学来实现。 在两个系统中协调基因表达和跨膜信号传导的大分子复合物， 依赖于高度合作的相互作用来响应光、氧化还原和化学环境。第一，细菌 趋化性，涉及原核细胞朝向外部刺激物的运动。趋化性是 理解跨膜通讯，细胞内信息传递和运动。重要的是， 许多引起霍乱、胃癌和莱姆病等疾病的人类病原体依赖于趋化性， 建立感染。趋化性背后的感觉器官，以下称为“化学体”， 惊人的灵敏度动态范围和基本的分子记忆在化学体，受体， 组氨酸激酶（CheA）和偶联蛋白组装成一个特定的结构，其细节只是 正在浮现该建议继续努力了解化学体组装，化学感受器构象 信号传导，并最终通过受体阵列的重组来调节CheA。化学体输出 调节着自然界完美的纳米机器--鞭毛马达。开关复合体的超微结构 将被定义为了解转矩产生，方向切换和响应， 化学体信号。第二个系统，真核生物钟，包括细胞自主计时 使新陈代谢与昼夜循环同步的装置。生物钟由转录-翻译 反馈环（TTFL），其中阻遏蛋白抑制其自身基因的转录激活因子。 光通过刺激直接撞击TTFL的光电传感器来夹带时钟相位。在人类中， 生物钟功能异常导致精神疾病（睡眠障碍，抑郁症，躁狂症），细胞生长失调 （癌症）和代谢缺陷（糖尿病和肥胖）。该项目提出了结构和机制 调查的关键阻遏物和光设置活动共同时钟在高等生物体。 将对真菌（粗糙脉孢菌）和苍蝇的昼夜节律蛋白进行生物物理研究 （黑腹果蝇）。这两种模式生物都提供了遗传系统和行为测定来探索 机械论观点的生物学意义。一套免费的技术，包括X射线 晶体学、小角X射线散射、光谱学、低温电子显微镜和脉冲偶极 ESR光谱（PDS）将被应用于实现这些目标。对于PDS，新的战略， 将开发和部署基于氮氧化物、黄素、核苷酸和金属离子的自旋探针。总的来说， 该计划旨在提供对细菌趋化性中的传感和反应的分子理解， 真核生物的昼夜节律通过生物物理方法的协同应用。