Phosphor-regulation of CAP1 Functions in Cell Adhesion and Migration

CAP1 在细胞粘附和迁移中的荧光调节功能

• 批准号: 9813335
• 负责人: Guolei Zhou
• 金额: $ 41.92万
• 依托单位: ARKANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9813335
• 关键词: Actins; Adhesions; Area; Arkansas; Biological Assay; Biological Process; Biomedical Research; Cancer Patient; Cell Adhesion; Cell Adhesion Molecules; Cell physiology; Cells; Cessation of life; Co-Immunoprecipitations; Complex; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinase 5; Cytoskeletal Proteins; Cytoskeleton; Data; Discipline; Embryonic Development; Environment; Eukaryota; Focal Adhesion Kinase 1; Future; Goals; Hela Cells; Human; Integrin-mediated Cell Adhesion Pathway; Knowledge; Laboratories; Lead; Life; Light; Link; Malignant Neoplasms; Mediating; Medicine; Microfilaments; Mission; Molecular; National Institute of General Medical Sciences; Neoplasm Metastasis; Outcome; Pathologic; Phosphorylation; Protein Dephosphorylation; Protein Isoforms; Proteins; Regulation; Role; Science; Signal Transduction; Site; Students; System; Talin; United States National Institutes of Health; Universities; Veins; Work; Wound Healing; Yeasts; base; cancer invasiveness; career; cell motility; cell type; cofilin; innovation; insight; migration; mutant; novel; response

PROJECT SUMMARY Cell migration is important for normal biological processes including embryonic development and wound healing, as well as pathological responses such as cancer metastasis. Cyclase-Associated Protein (CAP) is conserved in eukaryotes as a versatile actin-regulating protein. Mammalian CAP1 regulates the actin cytoskeleton and cell migration, and is also implicated in the invasiveness of human cancers. However, roles in cell migration and cancer invasiveness appear to be context-dependent, with insufficiently defined mechanisms. In that vein, our finding of a novel function for CAP1 in cell adhesion provided a mechanism that likely underlies the distinct roles for CAP1 in cell migration and cancer invasiveness. Moreover, we identified the very first regulation mechanism for CAP, where phosphorylation on both residues of the S307/S309 tandem site controls CAP1 association with cofilin and actin. Our latest preliminary studies provide compelling evidence that transient phosphorylation is required for CAP1 to promote cellular actin filament turnover and regulate integrin-mediated cell adhesion. Moreover, the Cyclin-Dependent Kinase 5 (CDK5) and cAMP antagonistically regulate S307/S309 phosphorylation on CAP1. However, critical gaps remain in molecular mechanisms underlying the phosphor-regulation of CAP1 by CDK5 and cAMP, and roles and mechanisms for this regulation in mediating the cell signals to control cell adhesion and migration. We propose this project to fill these critical gaps. Our long-term goal is to understand how cell signaling controls the actin cytoskeleton, cell migration and cancer invasiveness. The overall hypothesis is that CDK5 and cAMP signals antagonistically regulate CAP1 phosphorylation to facilitate spatial-temporally controlled phosphorylation, which is believed to be critical in regulating actin dynamics, cell adhesion and migration. We propose to pursue three specific aims: (1) further determine molecular mechanisms underlying the phosphorylation of CAP1 by CDK5 and dephosphorylation induced by cAMP, respectively; (2) ascertain roles for the phosphor-regulation of CAP1 in mediating CDK5 and cAMP signals to control cell adhesion and migration; (3) determine if S307/S309 phosphorylation regulates CAP1 association with FAK, talin or Rap1, which may underlie the phosphor- regulation of CAP1 function in cell adhesion. The proposed project is significant because successful completion of the specific aims will help us much better understand roles and mechanisms of the phosphor- regulation of the cytoskeletal protein CAP1, which links CAP1 phosphor-regulatory signals to the actin- dependent functions in adhesion and migration. The expected findings will not only greatly extend our knowledge on the regulation of CAP1 cell functions, but may also lead to strategies targeting CAP1 and/or its regulatory cell signals for suppressing the invasive cycle of cancer. This project is highly innovative, because it represents a further departure from the status quo, namely the already paradigm-shifting ongoing studies in our laboratory over the last several years, particularly that on the phosphor-regulation of CAP1.

项目总结 细胞迁移对正常的生物过程很重要，包括胚胎发育和 伤口愈合，以及癌症转移等病理反应。环化酶相关蛋白 (CAP)在真核生物中作为一种多功能的肌动蛋白调节蛋白而被保守。哺乳动物的Cap1蛋白调节肌动蛋白 细胞骨架和细胞迁移，也与人类癌症的侵袭性有关。但是，角色在 细胞迁移和癌症侵袭性似乎是依赖于上下文的，但定义不够充分 机制。在这方面，我们对cap1在细胞黏附中的新功能的发现提供了一种机制，即 这可能是Cap1在细胞迁移和癌症侵袭中的不同作用的基础。此外，我们还确定了 CAP的第一个调节机制，其中S307/S309的两个残基上的磷酸化 串联位点控制着cap1与粘连蛋白和肌动蛋白的关联。我们最新的初步研究提供了令人信服的 有证据表明，Cap1需要瞬时磷酸化才能促进细胞肌动蛋白细丝的周转 调节整合素介导的细胞黏附。此外，细胞周期蛋白依赖性激酶5(CDK5)和cAMP 拮抗调节CAP1上S307/S309的磷酸化。然而，关键的缺口仍然存在于分子水平 CDK5和cAMP对Cap1的磷调节机制及其作用和机制 这种调控在介导细胞信号的同时控制细胞的黏附和迁移。我们建议这个项目来填补 这些关键差距。我们的长期目标是了解细胞信号如何控制肌动蛋白细胞骨架 移民和癌症侵袭性。总体假设是CDK5和cAMP信号相互拮抗 调节cap1的磷酸化以促进时空受控的磷酸化，这被认为是 在调节肌动蛋白动态、细胞黏附和迁移方面起关键作用。我们建议达致三个具体目标： (1)进一步确定CDK5磷酸化cap1的分子机制和 CAMP诱导的去磷酸化；(2)确定Cap1在细胞内的磷酸化调节作用。 介导CDK5和cAMP信号调控细胞黏附和迁移；(3)确定S307/S309 磷酸化调节Cap1与FAK、talin或Rap1的关联，这可能是磷光体- CAP1在细胞黏附中的调节作用。拟议的项目意义重大，因为它成功了 具体目标的完成将有助于我们更好地理解荧光粉的作用和机理-- 细胞骨架蛋白Cap1的调节，它将Cap1磷酸化调节信号与肌动蛋白联系在一起。 在黏附和迁移中的依赖功能。预期的发现不仅将极大地扩展我们的 关于调节Cap1细胞功能的知识，但也可能导致针对Cap1和/或其 抑制癌症侵袭周期的调控细胞信号。这个项目很有创新性，因为它 代表着进一步背离现状，即#年已经在改变范式的正在进行的研究 我们的实验室在过去的几年里，特别是在CAP1的磷调节方面的研究。