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.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 背景：O-GlcNAc(b-N-乙酰氨基葡萄糖)是一种由N-乙酰氨基葡萄糖残基连接到数百种细胞质和核蛋白上的丝氨酸或苏氨酸的蛋白质修饰。蛋白质O-GlcNAc转移酶(OGT)催化糖部分的加成。在有丝分裂期间，OGT定位于离散的结构，OGT水平的升高会产生非整倍体，这是癌症的一个标志。 理论基础：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体这一假设驱动的。这项研究背后的理论基础是，一旦了解了OGT有丝分裂靶向背后的机制，这些相互作用就可以通过新的高度特异的药理学方法来操纵，从而产生癌症的治疗方法。 研究问题：目前尚不清楚OGT蛋白表达增加是如何导致非整倍体的。如果OGT在M期修饰了数百种蛋白质，那么OGT是如何针对特定底物的？非整倍体的表型是由于酶的活性还是OGT作为有丝分裂蛋白的支架？Aurora Kinase B如何与OGT相互作用？ 研究设计：我们将使用活细胞成像来研究OGT通过有丝分裂定位的动力学，使用质谱学来鉴定新的相互作用蛋白，最后定位OGT与其已知结合伙伴Aurora Kinase B之间的相互作用。 结果指标：这一目标的成功完成也将提供关于OGT定位的结构的信息，识别新的OGT靶向蛋白，最后我们将绘制OGT和Aurora Kinase B之间的相互作用结构域，并确定这种相互作用的中断如何改变M期进程。