Mechanisms of assembly and inheritance of yeast septin-containing structures

含有酵母septin的结构的组装和遗传机制

• 批准号: 7572147
• 负责人: MICHAEL A MCMURRAY
• 金额: $ 9万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7572147
• 关键词: Affinity; Attention; Award; Biological Assay; Biological Models; Cell division; Cell physiology; Cell-Free System; Cells; Cellular Structures; Complex; Deposition; Detection; Development; Disease; Electrons; Employee Strikes; Equilibrium; Eukaryota; Filament; Fluorescence; Foundations; Genes; Higher Order Chromatin Structure; Human; In Vitro; Individual; Kinetics; Label; Learning; Location; Malignant Neoplasms; Mediating; Modeling; Modification; Molecular; Molecular Chaperones; Monitor; Mothers; Mutation; Myosin Type II; Neck; Neurons; Organism; Physiological; Post-Translational Protein Processing; Principal Investigator; Proteins; Recombinants; Research; Research Personnel; Resolution; Role; Saccharomyces cerevisiae; Saccharomycetales; Staining method; Stains; Structure; System; Technology; Testing; Variant; Work; Yeasts; base; cell type; hereditary neuropathy; human disease; in vivo; insight; macromolecular assembly; novel; research study; stoichiometry; yeast protein

DESCRIPTION (provided by applicant): Septins are widely conserved among eukaryotes and implicated in a variety of cellular processes, although their precise molecular functions remain unknown. In every organism so far examined, multiple septin proteins are found in hetero-oligomeric complexes, the proper organization of which is required for their function. Mutation or misregulation that upsets the stoichiometry of septin hetero-oligomers is a common feature of septin-associated human diseases, which include cancer and hereditary neuropathies. Septin-containing cellular structures in dividing and differentiating cells are dynamic, undergoing abrupt changes in organization in a temporally and spatially regulated manner. The recent high-resolution structures of a human septin hexamer and a budding yeast octamer reveal the striking similarity of the constituent subunits and the interfaces by which they assemble. It is not known how assemblies with the proper arrangement of septin subunits are built in the cell, or how they are reorganized during cycles of proliferation and development. In this regard, the yeast septins represent a simple model system with which to identify cellular mechanisms regulating the structural organization of these multi- subunit macromolecular assemblies. Experiments supported by this award will examine how individual septin proteins are used in different assemblies, and how covalent modifications and septin-associated factors influence the kinetics of higher-order septin assembly. These studies adapt SNAP-tag technology for labeling of yeast proteins, allowing individual septins to be marked with a variety of functional tags and tracked through cell division and differentiation. Yeast septins are thought to form filaments at the mother-bud neck, but there has been no direct test of whether and how septins are organized in these structures. Experiments directly testing a model based on the structure of septin octamers aim to resolve this long-standing question. Finally, the same SNAP-Tag approach used to study septin dynamics will be applied to another dynamic assembly (myosin II-based filaments), to determine the generality of the new mechanistic insights revealed by the proposed research. Together, this work will lay the foundation for a long- term direction of study as an independent investigator. RELEVANCE: Although their molecular functions are not understood, it appears that the proper organization of different septin proteinss into multisubunit assemblies is critical. Indeed, a number of diseases (ranging from neuronal disorders to cancer) involve mutations in or misregulation of septin genes in ways that upset the balance of septin assembly. This work will identify the ways that yeast cells build and maintain their septin complexes.

描述（由申请人提供）：胞裂蛋白在真核生物中广泛保守，并参与多种细胞过程，尽管其精确的分子功能仍然未知。在迄今为止检查的每种生物体中，在异源寡聚复合物中发现了多种septin蛋白，其功能需要适当的组织。扰乱septin异源寡聚体的化学计量的突变或失调是septin相关人类疾病的共同特征，所述疾病包括癌症和遗传性神经病。分裂和分化细胞中的含隔膜的细胞结构是动态的，以时间和空间调节的方式经历组织的突然变化。最近的高分辨率结构的人septin六聚体和芽殖酵母八聚体揭示了惊人的相似性的组成亚基和接口，他们组装。目前还不知道如何组装与septin亚基的正确安排是建立在细胞中，或如何在增殖和发展的周期中重组。在这方面，酵母胞菌素代表了一个简单的模型系统，用它来鉴定调节这些多亚基大分子组装体的结构组织的细胞机制。该奖项支持的实验将研究单个Septin蛋白如何用于不同的组装，以及共价修饰和Septin相关因素如何影响高阶Septin组装的动力学。这些研究采用SNAP标签技术来标记酵母蛋白，允许用各种功能标签标记单个septins，并通过细胞分裂和分化进行跟踪。酵母细胞分裂素被认为在母芽颈形成细丝，但没有直接测试细胞分裂素是否以及如何在这些结构中组织。直接测试基于septin八聚体结构的模型的实验旨在解决这个长期存在的问题。最后，用于研究septin动力学的相同SNAP-Tag方法将应用于另一种动态组装（基于肌球蛋白II的细丝），以确定拟议研究揭示的新机制见解的一般性。总之，这项工作将奠定基础，为长期的研究方向作为一个独立的调查员。 相关性：虽然它们的分子功能还不清楚，但似乎不同septin蛋白质正确组织成多亚基组装是至关重要的。事实上，许多疾病（从神经元疾病到癌症）都涉及Septin基因的突变或失调，从而破坏了Septin组装的平衡。这项工作将确定酵母细胞建立和维持其septin复合物的方式。