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.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 背景：O-GlcNAc（b-N-乙酰葡糖胺）是一种蛋白质修饰，由连接到数百种细胞质和核蛋白上的丝氨酸或苏氨酸的 N-乙酰葡糖胺残基组成。 蛋白质 O-GlcNAc 转移酶 (OGT) 催化糖部分的添加。 在有丝分裂期间，OGT 定位于离散结构，OGT 水平升高会产生非整倍性，这是癌症的标志。 理由：这项研究的假设是 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。 这项研究背后的基本原理是，一旦了解了 OGT 有丝分裂靶向背后的机制，就可以通过新的、高度特异性的药理学方法来操纵这些相互作用，从而治疗癌症。 研究问题：目前尚不清楚的是 OGT 蛋白表达增加如何导致非整倍性。 如果 OGT 在 M 期修饰数百种蛋白质，那么 OGT 如何靶向特定底物？非整倍体表型是由酶活性引起的还是 OGT 充当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 OGT 通过有丝分裂定位的动力学，使用质谱法来识别新型相互作用蛋白，最后绘制 OGT 与其已知结合伴侣 Aurora 激酶 B 之间的相互作用图谱。 结果衡量：成功完成这一目标将提供有关 OGT 定位结构的信息，识别新的 OGT 靶向蛋白，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 期进展。