Regulation of Lens Fiber Cell Structure and Function by Post Translational Modification of Intermediate Filaments

通过中间丝的翻译后修饰调节晶状体纤维细胞结构和功能

基本信息

批准号：
9900012
负责人：
PAUL Gillespie FITZGERALD
金额：
$ 39.25万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-03-01 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9900012
关键词：
Affect Age Aging Antibodies Appearance Atomic Force Microscopy Biological Models Biology Caliber Caspase Cell Differentiation process Cell Maturation Cell physiology Cells Cellular Structures Chronology Clustered Regularly Interspaced Short Palindromic Repeats Crystalline Lens Data Electron Microscopy Event Genes Genetic Engineering Genetic Transcription Human Intermediate Filament Proteins Intermediate Filaments Lens Fiber Life Light Lipids Maps Masks Mass Spectrum Analysis Mechanics Mediating Mitosis Modeling Modification Molecular Mus Mutation Neonatal Optics Organelles Organism Phenotype Phosphorylation Phosphorylation Site Post-Translational Modification Site Post-Translational Protein Processing Process Protein Biosynthesis Proteins Publishing Regulation Reporting Resistance Resolution Rest Series Site Structure Surface Testing Thick Time Tissues Translating Translations Work Zebrafish cell type experimental study fiber cell lens mechanical properties mouse model mutant programs protein function temporal measurement

项目摘要

Project Summary/Abstract The ocular lens is an excellent model in which to study cellular and molecular aging. The lens is arranged as a series of concentric shells each 1 fiber cell thick. Lenses grow throughout life by addition of new shells to existing shells. Because no cells are lost with age, the shells are arranged in exact chronological order by birthdate, with the oldest at the center, and youngest at the surface. In humans this is about 2,500 concentric shells. Even more remarkable, fiber cells retain all their membranous organelles only until they complete formation of a new shell, at which point they are destroyed. In human lenses, only the outer 100 or so of the 2,500 layers has organelles. The other 2,400 layers are incapable of protein synthesis, and live the lifetime of the organism without protein replacement. Despite the inability to synthesize new protein, fiber cells undergo dramatic structural changes well past the point at which they have lost organelles. On the basis of preliminary data, and previously published work, we hypothesize that these structural changes are powered by a progression of post translational modifications (PTMs) to lens-specific intermediate filament (IF) proteins. We hypothesize that these PTMs change protein function which then orchestrates structural change. The field of lens biology is revealing a large number of similar processes that progress in cells lacking biosynthetic potential. We believe this proposal can directly test the cause (PTM) and effect (structural change) for at least one set of these changes, and do so with cell‐level resolution. Successful completion of this proposal will establish a mechanism by which cells can effect a progression of predictable structural changes, over time, in the absence of protein synthesis. To do this, we will identify PTMs, localize them with respect to structural change, then test the effect of the PTM directly by using CRISPR to genetically modify zebrafish IF PTM sites, and then translate key observation from zebrafish, into a mouse model.

项目摘要/摘要眼镜是研究细胞和分子衰老的绝佳模型。镜头是每个1个纤维细胞厚。镜片在一生中生长在现有外壳中添加新外壳。因为没有随着年龄的年龄而丢失的细胞，所以壳是按照生日按准确的按时间顺序排列，中心最古老，最年轻表面。在人类中，这大约是2500个同心壳。更引人注目的是纤维细胞仅保留所有膜细胞器，直到完成新外壳的形成为止他们被摧毁了。在人镜片中，只有2500层的外部100 有细胞器。其他2,400层是蛋白质合成的能力，并过着寿命无蛋白质替代的生物体。尽管无法合成新蛋白质，但纤维细胞经历了急剧的结构变化，远远超过了它们失去细胞器的点。根据初步数据和以前发表的工作，我们假设这些结构变化是由后翻译修饰（PTM）的进展为动力的透镜特异性中间丝（IF）蛋白。我们假设这些PTM改变了蛋白质功能，然后策划结构变化。镜头生物学领域正在揭示在缺乏生物合成潜力的细胞中进展的大量类似过程。我们相信该建议可以直接测试原因（PTM）和效果（结构性变化）至少这些更改的一组，并通过细胞级分辨率进行。成功完成建议将建立一种机制，通过该机制，细胞可以影响可预测的进展在没有蛋白质合成的情况下，结构变化，随着时间的流逝。为此，我们将确定 PTMS，将它们定位在结构变化方面，然后直接测试PTM的效果如果PTM站点，使用CRISPR通常修改斑马鱼，然后翻译关键观察从斑马鱼到鼠标模型。