STRUCTURE OF KINETOCHORE-MICROTUBULE INTERACTIONS

动粒-微管相互作用的结构

• 批准号: 8362526
• 负责人: MARY MORPHEW
• 金额: $ 1.06万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362526
• 关键词: 3-Dimensional; Binding; Cell physiology; Cells; Cellular Structures; Characteristics; Chromosomes; Flare; Freezing; Funding; Grant; Heterochromatin; In Vitro; Kinetochores; Methods; Microtubules; Mitotic; Mitotic spindle; Morphology; National Center for Research Resources; Principal Investigator; Research; Research Infrastructure; Resources; Slice; Source; Staging; Structure; United States National Institutes of Health; cost; in vivo; novel; reconstruction; tomography; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We are using cryofixed, freeze-substituted cells to study the morphology of kinetochores and their associated microtubule (MT) in mitotic cells. We have developed successful methods for preparing well-frozen mitotic cells (Morphew and McIntosh, J. Micros 212:21-25). 3-D reconstructions of kinetochores from several species have been obtained using dual-axis tomography. The structures of kinetochore-associated MT ends have been oriented and extracted using the "slicer" tool in IMOD. The walls of most kinetochore MTs flare outward at their plus ends, a morphology characteristic of disassembling MTs in vitro (Mandelkow et al., JCB 114:977, 1991). The degree of flaring has been quantified and found to be similarly distributed at all mitotic stages. These findings suggest that flaring in vivo is not simply indicative of disassembly, but may reflect a different dynamic state that can be controlled by other cellular processes. The regions around kinetochore MTs contain slender fibrils that appear to connect the MT walls in the centromeric heterochromatin. These findings suggest a novel mechanism for the binding of chromosomes to the mitotic spindle.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 我们正在使用冷冻的，冻结的细胞来研究有丝分裂细胞中动力学及其相关微管（MT）的形态。我们开发了制备良好的有丝分裂细胞的成功方法（Morphew和McIntosh，J。Micros 212：21-25）。使用双轴断层扫描获得了来自几种种类的动力学的3-D重建。 使用IMOD中的“切片机”工具将与动力学相关的MT末端的结构定向和提取。 大多数动力学MTS向外闪烁的墙壁的壁是其在其体外拆卸MTS的形态特征（Mandelkow等，JCB 114：977，1991）。耀斑的程度已被定量，并在所有有丝分裂阶段都类似地分布。 这些发现表明，体内燃烧不仅表明拆卸，而且可能反映了可以通过其他细胞过程控制的不同动态状态。 动力学MT周围的区域包含纤细的原纤维，这些原纤维似乎连接了丝粒异染色质中的MT壁。 这些发现提出了一种新的机制，可以使染色体与有丝分裂纺锤体结合。