Molecular Mechanics and Regulation of Centrosomes

中心体的分子力学和调控

• 批准号: 8644562
• 负责人: Erik Christian Yusko
• 金额: $ 5.15万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644562
• 关键词: Affect; Alanine; Anaphase; Aspartate; Binding; Biochemical; Cell Cycle; Cell Nucleus; Cell Survival; Cell division; Cells; Cellular biology; Centrosome; Chromosome Segregation; Chromosomes; Cytokinesis; Enzymes; Face; Feedback; Fluorescence Microscopy; Genomic Instability; In Vitro; Individual; Lasers; Lead; Light; Malignant Neoplasms; Measures; Mechanics; Metaphase; Methods; Microtubules; Mitotic spindle; Molecular; Mutation; Nuclear; Organism; Pathway interactions; Phosphorylation; Phosphorylation Site; Positioning Attribute; Property; Prophase; Proteins; Regulation; Role; Rupture; Site; Staging; Structure; Supporting Cell; Techniques; Testing; Tubulin; Weight-Bearing state; Work; Yeasts; base; cell motility; fly; human disease; in vivo; molecular mechanics; mutant; public health relevance; reconstitution; research study; single molecule; tumor progression

DESCRIPTION (provided by applicant): Centrosomes are vital mechanical hubs in cells that sustain load-bearing attachments to microtubules and orchestrate arrays of microtubules to support cell motility, nuclear positioning, mitotic spindle assembly, and chromosome segregation. Mutations affecting the ability of centrosomes to sustain or regulate attachments of microtubules are correlated with an increase in genome instability and progression of cancers. Despite this important role and the considerable forces that centrosome-microtubule interactions are thought to sustain in vivo, microtubule attachments to centrosomes have never been mechanically investigated in any organism. Consequently, the extent to which these interactions may be mechanically regulated has also never been investigated. In a collaborative project with the Asbury, Davis, and Agard labs, this work aims to reveal the molecular interactions by which centrosomes sustain, sense, and respond to forces transmitted by microtubules. Centrosome-microtubule interactions will be reconstituted in vitro and individually interrogated using a combination of single molecule laser trapping, total internal reflection fluorescence (TIRF) microscopy, and molecular cell biology techniques. Through the combination of biophysical expertise in the Asbury lab with the biochemical and structural expertise of the Davis and Agard labs, this proposal will determine the role of specific molecular components in sustaining and regulating attachments of microtubules to centrosomes more precisely and directly than has ever been done before. Revealing how centrosomes sustain, sense, and respond to mechanical forces will shed light on how these pathways fail in human diseases such as cancer and may ultimately lead to the identification of new chemotherapeutic targets.

描述（由申请人提供）：中心体是细胞中至关重要的机械枢纽，可维持与微管的承重连接，并协调微管阵列，以支持细胞运动、核定位、有丝分裂纺锤体组装和染色体分离。影响中心体维持或调节微管附着能力的突变与基因组不稳定性增加和癌症进展相关。尽管这一重要作用和相当大的力量，中心体微管相互作用被认为是在体内维持，微管附件的中心体从来没有在任何生物体进行机械研究。因此，在何种程度上，这些相互作用可能是机械调节也从未被调查。在与阿斯伯里、戴维斯和阿加德实验室的一个合作项目中，这项工作旨在揭示中心体维持、感知和响应微管传递的力的分子相互作用。中心体-微管相互作用将在体外重建，并使用单分子激光捕获、全内反射荧光（TIRF）显微镜和分子细胞生物学技术的组合进行单独询问。通过结合阿斯伯里实验室的生物物理学专业知识和戴维斯和阿加德实验室的生物化学和结构专业知识，该提案将确定特定分子组分在维持和调节微管与中心体的连接中的作用，比以往任何时候都更精确和直接。揭示中心体如何维持，感知和响应机械力将揭示这些途径如何在人类疾病（如癌症）中失败，并可能最终导致新的化疗靶点的鉴定。