Targeting and Regulation of O-GlcNAc Transferase at M Phase

M 期 O-GlcNAc 转移酶的靶向和调节

基本信息

批准号：
8480366
负责人：
Chad Eric Slawson
金额：
$ 22.65万
依托单位：
UNIVERSITY OF KANSAS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8480366
关键词：
Acetylglucosamine Affinity Amino Acids Aneuploidy Binding Biological Cell Cycle Progression Cells Centrosome Complex Coupled Cytokinesis Cytoplasmic Protein Data Development Disease Environment Enzymes Eukaryotic Cell Goals Grant Growth Holoenzymes Hormones Image Kinetics Laboratories Lead Malignant Neoplasms Mass Spectrum Analysis Measures Mitosis Mitotic Mitotic spindle Modification Molecular Nuclear Proteins Nutrient O-GlcNAc transferase Oligosaccharides Outcome Phenotype Phosphotransferases Post-Translational Protein Processing Proteins Proteomics Recovery Regulation Research Role Serine Signal Transduction Solid Neoplasm Source Stress Structure Techniques Testing Threonine Time UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase aurora B kinase base cancer therapy domain mapping extracellular gain of function insight novel poly-N-acetyl glucosamine protein expression sugar tumor progression

项目摘要

O-GlcNAc is the covalent modification of proteins at serine/threonine amino acids by the sugar moiety Nacetylglucosamine. The O-GlcNAc modification is not elongated to more complex oligosaccharide structures and is found only on nuclear and cytoplasmic proteins. The modification is highly dynamic and responds to signals of the extracellular environment such hormones, stress, and nutrients. Previously, I demonstrated that 0-GlcNAc is critical for the proper progression of the cell cycle in eukaryotic cells; gain of function of either 0-GlcNAc transferase (OGT), the enzyme which adds the modification, or 0-GlcNAcase (OGA), the enzyme which removes the modification, causes mitotic exit delays. Furthermore, OGT and OGA can be found in a signaling complex with mitotic kinases such as Aurora Kinase B. The goal of this proposal is to investigate the interactions of OGT with mitotic structures such as the spindle and with mitotic proteins. We hypothesize that OGT forms complexes with various proteins during M phase progression that then targets OGT the spindle and midbody. I plan to use multiple techniques such as using FRAP (Fluorescent Recovery after Photo-bleaching) on GFP-OGT to measure the kinetics of OGT at spindle/midbody, quantitative proteomics to identify OGT targeting proteins, and finally identifying the function of Aurora Kinase B in targeting OGT to mitotic substrates and structures. Our expected contribution is significant because we expect to find insight into the regulation of OGT by mitotic targeting proteins and how these interactions are uncoupled in diseases such as cancer.

O-GLCNAC是通过糖部分naceTylyl葡萄糖胺在丝氨酸/苏氨酸氨基酸上对蛋白质的共价修饰。 O-GLCNAC的修饰并未延长到更复杂的寡糖结构，仅在核和细胞质蛋白上发现。这种修饰是高度动态的，并响应细胞外环境的信号，例如激素，压力和养分。以前，我证明了0-GLCNAC对于真核细胞中细胞周期的正确进展至关重要。功能 0-GLCNAC转移酶（OGT），添加修饰的酶或0-GLCNACASE（OGA），即去除修饰的酶，会导致有丝分裂出口延迟。此外，OGT和OGA可以在与Aurora激酶等有丝分裂激酶的信号传导复合物中找到。该建议的目的是研究OGT与有丝分裂结构（例如纺锤体和有丝分裂蛋白）的相互作用。我们假设OGT在M相进展过程中与各种蛋白质形成复合物，然后将OGT靶向主轴和中体。 I plan to use multiple techniques such as using FRAP (Fluorescent Recovery after Photo-bleaching) on GFP-OGT to measure the kinetics of OGT at spindle/midbody, quantitative proteomics to identify OGT targeting proteins, and finally identifying the function of Aurora Kinase B in targeting OGT to mitotic substrates and structures.我们的预期贡献很重要，因为我们期望通过有丝分裂靶向蛋白以及如何在癌症等疾病中发现这些相互作用如何对OGT的调节进行了解。