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Phosphatase-dependent regulation of desmosome intercellular junctions

桥粒细胞间连接的磷酸酶依赖性调节

基本信息

批准号：
10677182
负责人：
Abbey Leigh Perl
金额：
$ 7.18万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10677182
关键词：
Acantholysis Address Adhesions Adhesives Affect Affinity Apical Binding C-terminal Cadherins Cell Adhesion Cell division Cell-Cell Adhesion Cells Coupled Couples Cutaneous Cytoskeleton DNA Sequence Alteration Data Defect Deficiency Diseases Deletion Mutation Desmosomes Development Development Plans Disease Disease model Ectopic Expression Epidermis Epidermolysis Bullosa Equilibrium Event Exhibits Experimental Models Exposure to Family Dasypodidae Family member Foundations Funding Future Genetic Goals Human Immune In Vitro Intercellular Junctions Intermediate Filaments Keratin Label Maintenance Mechanical Stress Mechanics Mediating Membrane Microscopy Modeling Pemphigus Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Point Mutation Post-Translational Protein Processing Process Property Protein Dephosphorylation Protein Phosphatase Inhibitor Protein phosphatase Proteins Proteomics Regulation Research Personnel Role Signal Transduction Skin Stratification Stress Stretching Syndrome Tail Techniques Testing Tissues Training Work arrhythmogenic cardiomyopathy career career development desmoplakin desmoplakin II endosulfine genetic approach insight keratinocyte mechanical force mechanical properties mechanotransduction molecular phenotype mouse model mutant novel preservation protective effect recruit research and development response skin barrier superresolution microscopy trafficking training opportunity

项目摘要

Project Summary Intercellular junctions and their cytoskeletal connections are essential for maintaining tissue stability and function. This is particularly true of the multi-layered epidermis as it is exposed to high levels of mechanical stress while maintaining an essential physical and immune barrier. Critical contributors to the epidermis' ability to maintain the epidermal barrier while simultaneously adjusting to stress are attachments between keratin intermediate filaments (IF) and intercellular junctions called desmosomes. Within desmosomes, a cytoskeletal linker protein called desmoplakin (DP) is responsible for anchoring the IF to the junctions. DP is ubiquitously expressed in all desmosome-forming cells and is regulated by post-translational modification (PTM) of its C-terminal tail domain to control DP-IF affinity. Consistent with the importance of this phospho-regulatory region of DP for desmosome function, genetic deletions of the C-terminal region cause multiple diseases associated with severe cardio- cutaneous and lethal epidermal barrier defects. Functionally, when the C-terminal motif is in its hypo- phosphorylated state, DP exhibits its highest affinity towards IF. In experimental models, the increased DP-IF association had protective effects in mature cell sheets by generating stronger, more stable desmosomes. Conversely, during desmosome assembly hypo-phosphorylated DP accumulates on the IF network, disrupting it's trafficking to desmosome junctions, and interfering with the formation and maturation of new cell-cell desmosome junctions. Given the strong molecular phenotypes associated with DP's hypo-phosphorylated form in vitro, a coordinated process must exist to regulate DP phosphorylation for proper desmosome function. We previously identified GSK3 as the kinase responsible for phosphorylating DP; however, the protein phosphatase responsible for negatively regulating DP phosphorylation was previously unknown. My preliminary data identified PP2A-B55 as capable of binding to and dephosphorylating DP's C-terminus. This proposal will test the hypothesis that PP2A-B55 regulates DP phosphorylation during the dynamic process of desmosome assembly and in response to mechanical stress to allow cells to respond to the changing properties and specialized functions of the stratified epidermis. Aim 1 will employ cutting-edge microscopy and proteomics- based approaches to identify the mechanism by which PP2A regulates DP during desmosome assembly. Aim 2 will establish how PP2A phospho-regulation of DP impacts 1) desmosome-dependent cell adhesion and 2) the desmosomal response to mechanical stress using novel mechano-biology techniques. The proposed work will provide important insight into how phosphatase signaling controls desmosome assembly and function to maintain the human epidermal barrier and, therefore, how dysregulated phosphatase signaling could contribute to barrier-related diseases. Together, the proposed research and career development plan will provide a springboard for my development as an independent investigator and planned K99/R00 application.

项目总结