Mechanisms of centrosome regulation

中心体调节机制

• 批准号: 9034018
• 负责人: Dorothy A Lerit
• 金额: $ 24.9万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9034018
• 关键词: Address; Advisory Committees; Affinity Chromatography; Area; Award; Biology; Cancer Etiology; Career Transition Award; Cell Culture Techniques; Cell Cycle; Cell division; Cells; Centrioles; Centrosome; Cilia; Clustered Regularly Interspaced Short Palindromic Repeats; Communication; Computer Simulation; Confocal Microscopy; Development; Development Plans; Disease; Drosophila genus; Educational workshop; Elements; Embryo; Embryonic Development; Eukaryotic Cell; Evaluation; Event; Extramural Activities; Feedback; Flagella; Fluorescence Recovery After Photobleaching; Functional disorder; Funding; G-substrate; Generations; Genetic Transcription; Genome; Growth; Hand; Image; Interphase; Label; MS2 coat protein; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Mentors; Messenger RNA; Methods; Microcephaly; Microscopy; Microtubule-Organizing Center; Mitosis; Mitotic; Mitotic spindle; Modeling; National Heart, Lung, and Blood Institute; Pathologic; Pattern; Phase; Post-Transcriptional Regulation; Principal Investigator; Proteins; RNA; RNA Interference; RNA Sequences; RNA-Binding Proteins; Recruitment Activity; Regulation; Reporter; Reporting; Research; Research Personnel; Resolution; Role; Science; Shapes; Sterility; Structure; Technical Expertise; Technology; Testing; Tissues; Training; Transcript; United States National Institutes of Health; Work; base; career; career development; career preparation; cell motility; cell type; ciliopathy; cilium biogenesis; daughter cell; experience; gamma Tubulin; genome editing; hands on instruction; insight; knock-down; member; migration; mutant; neglect; nerve stem cell; neuroblast; novel; pericentrin; polarized cell; protein complex; protein function; research and development; scaffold; spatiotemporal; stem cell division; trafficking; tumor progression

 DESCRIPTION (provided by applicant): Centrosome dysfunction is the leading cause of microcephaly and is also associated with sterility, ciliopathy, and cancer. As the microtubule organizing centers of most eukaryotic cells, centrosomes build the bipolar mitotic spindle that segregates the duplicated genome into two daughter cells during cell division. In addition, centrosomes orchestrate numerous critical tasks during interphase, such as cell polarization, ciliogenesis, cell migration, and intracellular trafficking. The proposed research seeks to uncover novel paradigms of centrosome regulation. The results from this work are likely to fundamentally reshape current models of how centrosome activity is controlled in healthy and diseased cells. While most centrosome studies focus on maturation, the recruitment of proteins to the centrosome at mitotic onset, how centrosomes are regulated during interphase is little understood. In Aim 1, I will determine how centrosome function is regulated during interphase by Pericentrin-like-protein. Further, I propose to investigate the unknown roles of mRNAs that were previously localized to centrosomes. Many of these transcripts encode regulators of centrosome activity. In Aim 2, I propose to visualize mRNAs that localize to centrosomes and to investigate RNA localization mechanisms. I hypothesize that mRNA localization is an efficient strategy to quickly and locally control centrosome activity. To test this hypothesis, I will elucidte the mechanism and function of mRNA localization to centrosomes in Aim 3. Investigating these unexplored paradigms of centrosome regulation is essential to understand how centrosome function is modulated throughout the cell cycle and deregulated in disease. The NHLBI K22 Career Transition Award will provide mentored research and career development training to support the successful completion of the proposed research and to facilitate my transition to career independence. This award proposal was developed to leverage my experience in mRNA localization with my current training in centrosome biology. As part of a comprehensive career development plan, the mentored phase will include hands-on instruction, formal workshops, mentoring opportunities, and science communication activities. New technical skills to be learned include fluorescence recovery after photobleaching, super resolution microscopy, cell culture, and CRISPR genome editing. To address deficiencies in these areas and to provide tailored career preparation, the mentors and expert members of an advisory committee, comprising both intramural and extramural investigators, will provide frequent discussion, feedback, and evaluation. With this information in hand, I will launch my independent career as a principal investigator. Findings from the proposed research will form the basis of an NIH R01 funding application. Further, the completion of the proposed work will provide a more comprehensive understanding of how centrosome activity is regulated, aspects of which may be deregulated in a disease setting.

 描述（由申请人提供）：中心体功能障碍是小头畸形的主要原因，也与不育、纤毛病变和癌症有关。中心体作为大多数真核细胞的微管组织中心，在细胞分裂过程中构建双极有丝分裂纺锤体，将复制的基因组分离成两个子细胞。此外，中心体在间期期间协调许多关键任务，如细胞极化，纤毛发生，细胞迁移和细胞内运输。这项拟议中的研究旨在揭示 中心体调节的新范例。这项工作的结果可能会从根本上重塑目前在健康和患病细胞中中心体活动如何控制的模型。 虽然大多数中心体的研究集中在成熟，有丝分裂开始时，中心体的蛋白质的招聘，如何在间期调节中心体是知之甚少。在目标1中，我将确定如何中心体的功能是在间期由Pericentrin样蛋白调节。此外，我建议调查未知的作用的mRNA，以前定位于中心体。这些转录本中的许多编码中心体活性的调节子。在目标2中，我建议可视化定位到中心体的mRNA，并研究RNA定位机制。我假设mRNA定位是一种快速和局部控制中心体活性的有效策略。为了验证这一假设，我将阐明mRNA定位到中心体的机制和功能在目标3。研究这些未被探索的中心体调控模式对于了解中心体功能如何在整个细胞周期中被调节以及在疾病中被解除调控是至关重要的。 NHLBI K22职业过渡奖将提供指导研究和职业发展培训，以支持成功完成拟议的研究，并促进我向职业独立的过渡。这个奖项的建议是利用我在mRNA定位的经验与我目前在中心体生物学的培训。作为全面职业发展计划的一部分，辅导阶段将包括实践指导、正式研讨会、辅导机会和科学交流活动。需要学习的新技术技能包括光漂白后的荧光恢复，超分辨率显微镜，细胞培养和CRISPR基因组编辑。为了解决这些领域的不足之处，并提供量身定制的职业准备，顾问委员会的导师和专家成员，包括校内和校外调查人员，将提供频繁的讨论，反馈和评估。有了这些信息，我将开始我作为首席研究员的独立职业生涯。拟议研究的结果将构成NIH R01资助申请的基础。此外，完成拟议的工作将提供一个更全面的了解中心体活动是如何调节的，其中一些方面可能会在疾病的设置解除管制。