Integration of regulatory networks and subcellular architecture to control the Caulobacter cell cycle

整合调控网络和亚细胞结构来控制柄杆菌细胞周期

• 批准号: 9281784
• 负责人: LUCILLE SHAPIRO
• 金额: $ 64.05万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9281784
• 关键词: Antibiotics; Architecture; Bacteria; Caulobacter; Caulobacter crescentus; Cell Cycle; Cell Differentiation process; Cell division; Cells; Chromosome Structures; Communicable Diseases; Complement; Development; Dimensions; Dissection; Epigenetic Process; Gene Expression; Generations; Genetic; Genetic Transcription; Genome; Goals; Growth; Light; Liposomes; Logic; Molecular; Organism; Pathway interactions; Research; Resistance; Signal Pathway; Signal Transduction; Signaling Protein; Solid; Structural Protein; System; Time; Translations; arctic environment; cell type; daughter cell; design; genetic regulatory protein; imaging modality; mutant; novel; optogenetics; programs; public health relevance; reconstitution; segregation; single molecule; spatiotemporal

 DESCRIPTION (provided by applicant): The fundamental basis for the generation of cellular diversity in all organisms is the asymmetric deployment of structural and regulatory proteins to the cell poles prior to cell division and the consequent differential readout of the genome in the two daughter cells. The bacterium Caulobacter crescentus provides an elegant system in which to decipher the complete molecular circuitry that controls the asymmetry that underlies cell differentiation. As Caulobacter moves through its cell cycle, cell differentiation is accompanied by the polar localization of distinct complements of phospho-signaling proteins. The goals of our research program turn on three important questions: 1- How does a polar matrix nanodomain function to dynamically re-wire polar phosphosignaling pathways to drive cell differentiation and asymmetric cell division? We are approaching this question through reconstitution of the polar environment using liposomes and microfabricated solid substrates, three dimensional superresolution imaging modalities and single molecule tracking in living cells, and the creation of optogenetic mutants that enable instantaneous light-induced reconfiguration of the cell pole composition, thereby allowing us to directly observe the consequences of re-wiring a spatially-restricted signaling cascade in a living cell. 2- How do cell-type specific signaling pathways beget cell type-specific gene expression? We are defining the exquisite complexity of the complete genetic circuitry that uses both transcriptional and translation control to drive cell cyce progression culminating in daughter cells of different cell fate. 3- How does chromosome organization, replication and segregation along the long axis of the cell serve as a timer of cell cycle-regulated transcription using epigenetic mechanisms? Our goal is to integrate these spatiotemporal regulatory paradigms to establish the logic that is applicable to the dissection of asymmetry in all living systems.

 描述(申请人提供)：在所有生物中产生细胞多样性的基本基础是结构和调节蛋白在细胞分裂前不对称地部署到细胞极点，从而在两个子细胞中差异读出基因组。新月形杆菌提供了一个优雅的系统，在这个系统中可以破译控制细胞分化的不对称性的完整分子电路。随着Caulbacter在其细胞周期中的移动，细胞分化伴随着不同的磷酸信号蛋白补体的极地定位。我们研究计划的目标集中在三个重要的问题上：1-极性基质纳米结构域如何发挥功能，动态重新连接极性磷光信号通路，以驱动细胞分化和不对称细胞分裂？我们正在通过使用脂质体和微型制造的固体底物重建极地环境，三维超分辨率成像模式和活细胞中的单分子跟踪，以及创造光遗传突变体来实现细胞极组成的瞬时光诱导重新配置，从而使我们能够直接观察在活细胞中重新连接空间受限的信号级联的后果。细胞类型特异性信号通路如何产生细胞类型特异性基因表达？我们正在定义完整遗传电路的精致复杂性，该电路使用转录和翻译控制来驱动细胞周期进展，最终导致不同细胞命运的子细胞。染色体的组织、复制和沿细胞长轴的分离如何利用表观遗传机制作为细胞周期调控转录的定时器？我们的目标是整合这些时空调控范式，以建立适用于剖析所有生命系统中的不对称性的逻辑。