The relationship between Rab11-endosomes and the centrosome during division

Rab11-核内体和中心体在分裂过程中的关系

• 批准号: 9892164
• 负责人: Heidi Hehnly
• 金额: $ 21万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-03 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9892164
• 关键词: 3-Dimensional; Acinus organ component; Address; Animal Model; Apical; Attention; Binding; Biosensor; Cell Culture Techniques; Cell Cycle; Cell division; Cell membrane; Cells; Centrioles; Centrosome; Cilia; Congenital Heart Defects; Cyst; Cytokinesis; DNA Sequence Alteration; Defect; Dependence; Development; Disease; Dissociation; Distal; Endosomes; Event; Fluorescence Resonance Energy Transfer; Genes; Guanosine Triphosphate Phosphohydrolases; Hair; Interphase; Kidney; Left; Link; Liver; Measures; Membrane; Microtubules; Mitotic; Mitotic spindle; Modeling; Molecular; Monitor; Monomeric GTP-Binding Proteins; Morphogenesis; Mothers; Oranges; Organ; Organelles; Pancreas; Pathway interactions; Patients; Play; Positioning Attribute; Process; Proteins; Recycling; Role; Signal Transduction; Site; Structure; Testing; Time; Tissues; Transport Vesicles; Tubular formation; Vertebrates; Vesicle; Vesicle Transport Pathway; Zebrafish; apical membrane; appendage; base; cell assembly; cell type; ciliopathy; cilium biogenesis; cilium motility; fluid flow; in vivo; kidney cell; kidney epithelial cell; kinetosome; live cell imaging; optogenetics; rab GTP-Binding Proteins; tool

SUMMARY STATEMENT: Ciliopathies comprise a group of disorders associated with genetic mutations encoding defective proteins, which result in either abnormal formation or function of cilia. Ciliopathies have been linked to defects in molecular pathways organized at the primary cilium's basal body/centrosome and the primary cilium itself. Patients suffering from ciliopathies can display defects in left/right asymmetry, congenital cardiac defects, and formation of cysts in multiple organs including liver, pancreas and kidney. In vertebrates, motile cilia located in an organ of asymmetry play an important role in left-right development. Evidence from model organisms, such as zebrafish organ of asymmetry (Kupffer's Vesicle, KV), indicates that conserved cilia-driven left-right flow establishes left- right signals to regulate target genes to control left/right asymmetry. This project addresses the question: How do ciliated cells develop into a functional polarized organ? We propose this occurs through a sequential process that starts with cell division, polarity formation, and finishes with cilia assembly. If a ciliated organ wants to expand its central lumen, cells need to re-enter the cell cycle and correctly position its mitotic spindle along the longest axis parallel to the expanding lumen. We found that both Rab11-associated endosomes and a centrosome protein, cenexin, regulate the positioning of the mitotic spindle and ciliogenesis (Hehnly and Doxsey, 2014; Hung et al., 2016), but whether they act together during these processes has not been examined. During interphase, Rab11-endosomes associate with mother centriole sub-distal appendages where cenexin resides (Hehnly et al., 2012). Rab11 and a second GTPase, Rab8, can directly interact with cenexin. In addition, a GTPase cascade has been identified between Rab11 and Rab8 during ciliogenesis and apical membrane initiation, but if it requires the centrosome or occurs during specific points of the cell cycle is unknown. We will test the overall hypothesis that anchoring Rab11 and/or Rab8 by the mother centriole appendage protein, cenexin, is required for spindle positioning, polarity formation and/or ciliogenesis. To test this we will determine whether the centrosome can modulate Rab11 and/or Rab8 activity (Aim 1) and determine when during the cell cycle Rab11 and Rab8 activity is required to regulate tissue morphogenesis (Aim 2). This will be accomplished by using a 3-D kidney epithelial cell culture model and an in vivo developmental model, zebrafish KV formation, to identify a framework for how polarity formation relates to the cell cycle.!

摘要：纤毛病包括一组与遗传相关的疾病 编码有缺陷的蛋白质的突变，导致纤毛的形成或功能异常。 纤毛病与初级纤毛基底组织的分子途径缺陷有关 体/中心体和初级纤毛本身。患有纤毛病的患者可能会表现出以下缺陷： 左/右不对称、先天性心脏缺陷以及多个器官囊肿的形成，包括 肝脏、胰腺和肾脏。在脊椎动物中，位于不对称器官中的活动纤毛起着 在左右发展中发挥着重要作用。来自模型生物的证据，例如斑马鱼的器官 不对称性（库普弗囊泡，KV），表明保守的纤毛驱动的左右流动建立了左- 右信号调节靶基因以控制左/右不对称。该项目解决了 问题：纤毛细胞如何发育成功能性极化器官？我们建议发生这种情况 通过一个连续的过程，从细胞分裂开始，极性形成，并以纤毛结束 集会。如果纤毛器官想要扩大其中央管腔，细胞需要重新进入细胞周期并 沿着平行于扩张管腔的最长轴正确定位其有丝分裂纺锤体。我们发现 Rab11 相关内体和中心体蛋白 cenexin 均调节 有丝分裂纺锤体和纤毛发生（Hehnly 和 Doxsey，2014；Hung 等人，2016），但它们是否起作用 尚未对这些过程中的一起进行检查。在间期期间，Rab11-内体 与 cenexin 所在的母体中心粒远端附件相关（Hehnly 等，2012）。 Rab11 和第二种 GTP 酶 Rab8 可以直接与 cenexin 相互作用。此外，GTPase 级联 在纤毛发生和顶膜起始过程中，Rab11 和 Rab8 之间已被鉴定，但如果 它需要中心体或发生在细胞周期的特定点尚不清楚。我们将测试 总体假设是通过母体中心粒附属蛋白锚定 Rab11 和/或 Rab8， cenexin 是纺锤体定位、极性形成和/或纤毛发生所必需的。为了测试这一点，我们将 确定中心体是否可以调节 Rab11 和/或 Rab8 活性（目标 1）并确定 当细胞周期期间需要 Rab11 和 Rab8 活性来调节组织形态发生时（目的 2）。这将通过使用 3D 肾上皮细胞培养模型和体内 发育模型，斑马鱼 KV 形成，以确定极性形成如何相关的框架 到细胞周期。！