TARGETING AND REGULATION OF O-GLCNAC TRANSFERASE DURING MITOSIS

有丝分裂期间 O-GLNAC 转移酶的靶向和调节

• 批准号: 8360686
• 负责人: Chad Eric Slawson
• 金额: $ 21.66万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360686
• 关键词: Acetylglucosamine; Aneuploidy; Binding; Cells; Complex; Cytoplasmic Protein; Development; Funding; Grant; Holoenzymes; Kinetics; Life; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mitosis; Mitotic; Molecular; National Center for Research Resources; Nuclear Proteins; O-GlcNAc transferase; Outcome Measure; Phenotype; Post-Translational Protein Processing; Principal Investigator; Proteins; Regulation; Research; Research Design; Research Infrastructure; Resources; Serine; Source; Structure; Threonine; United States National Institutes of Health; aurora B kinase; cancer therapy; cellular imaging; cost; novel; protein expression; scaffold; sugar

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Background: O-GlcNAc (b-N-acetylglucosamine) is a protein modification consisting of an N-acetylglucosamine residue attached to serines or threonines on hundreds of cytoplasmic and nuclear proteins. The protein O-GlcNAc transferase (OGT) catalyzes the addition of the sugar moiety. During mitosis, OGT localizes to discrete structures, and elevated levels of OGT produce aneuploidy, a hallmark of cancer. Rationale: This research is driven by the hypothesis that OGT forms unique holoenzyme complexes with specific proteins during mitosis. The rationale behind this research is that once the mechanism behind mitotic targeting of OGT is understood, these interactions can be manipulated by new and highly specific pharmacological approaches resulting in treatments for cancer. Study Questions: What is unclear is how increased OGT protein expression causes aneuploidy. If OGT modifies hundreds of proteins at M phase, then how is OGT targeted to specific substrates? Is the aneuploidy phenotype due to enzymatic activity or does OGT act as a scaffold for mitotic proteins? How does Aurora Kinase B interact with OGT? Study Design: We will employ live cell imaging to study the kinetics of OGT localization through mitosis, use mass spectrometry to identify novel interacting proteins, and lastly map the interaction between OGT and its known binding partner Aurora Kinase B. Outcome Measures: Successful completion of the this aim will provide information on the structures OGT localizes too, identify novel OGT targeting proteins, and finally we will map the interacting domains between OGT and Aurora Kinase B and determine how disruption of this interaction alters M phase progression.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 背景：O-GLCNAC（B-N-乙酰葡萄糖）是一种蛋白质修饰，该蛋白质由数百种细胞质和苏氨酸连接到数百种细胞质和核蛋白上的N-乙酰基葡萄糖残基组成。 蛋白O-GLCNAC转移酶（OGT）催化糖部分的添加。 在有丝分裂期间，OGT本地化为离散结构，OGT水平升高会产生非整倍性，这是癌症的标志。 理由：这项研究是由OGT在有丝分裂过程中与特定蛋白质形成独特的全酶复合物的假设所驱动的。 这项研究背后的理由是，一旦了解了OGT有丝分裂靶向的机制，这些相互作用就可以通过新的且高度特定的药理方法来操纵，从而导致癌症治疗。 研究问题：尚不清楚的是OGT蛋白表达增加如何引起非整倍性。 如果OGT在M期修饰数百种蛋白质，那么OGT如何针对特定的底物？是由于酶活性引起的非整倍性表型还是OGT是有丝分裂蛋白的支架？ Aurora激酶B如何与OGT相互作用？ 研究设计：我们将采用活细胞成像来研究通过有丝分裂的OGT定位动力学，使用质谱来鉴定新型相互作用蛋白，最后绘制OGT与其已知结合伴侣Aurora激酶B之间的相互作用。 结果指标：成功完成此目标将提供有关OGT结构的信息，也可以识别靶向蛋白质的新型OGT，最后我们将绘制OGT和Aurora激酶B之间的相互作用域，并确定这种相互作用的破坏如何改变M相位的进展。