SAXS ON TRANSCRIPTION FACTOR IIF ASSOCIAT C-TERMINAL REPEAT DOMAIN PHOSPHATASE

转录因子 IIF 相关 C 末端重复结构域磷酸酶上的 SAXS

• 批准号: 8363569
• 负责人: Neela H. Yennawar
• 金额: $ 0.55万
• 依托单位: CORNELL UNIVERSITY
• 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 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363569
• 关键词: Adopted; Binding; C-terminal; Chemicals; Data; Data Analyses; Disease; Equilibrium; Funding; General Transcription Factors; Grant; Hydrogen Bonding; L-Iditol 2-Dehydrogenase; Liver; Molecular Conformation; NMR Spectroscopy; National Center for Research Resources; Peptides; Phosphoric Monoester Hydrolases; Principal Investigator; RNA Polymerase II; Research; Research Infrastructure; Resources; Shapes; Sheep; Solutions; Source; Spectrum Analysis; Structure; Terminal Repeat Sequences; United States National Institutes of Health; carboxy-terminal domain phosphatase; cost; interest; transcription factor; transcription termination

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. Project 1 - Termination of transcription by RNA polymerase II is promoted by an interaction between the general transcription factor IIF (TFIIF) and the TFIIF-associating C-terminal repeat domain phosphatase (FCP1). The acidic C-terminal region of FCP1 is disordered in the free state, as indicated by NMR and CD spectroscopy, but adopts an ¿¿-helical conformation upon binding to the heavy chain of TFIIF. We have recently utilized 13C-detected NMR spectroscopy to generate comprehensive NMR chemical shifts of free ctFCP1 (residues 879-961) and secondary shift analysis of these data suggest that the equilibrium ensemble is biased towards ¿¿-helical conformation in residues 945-960 ¿¿" the TFIIF associating peptide ¿¿" despite the lack of persistent helical structure indicated by the CD results. We aim to gain further constraint on the overall hydrodynamic shape of the ctFCP1 solution ensemble through acquisition of SAXS data on the same 879-961 construct. Project 2 - We have recently solved the crystal structure of sheep liver sorbitol dehydrogenase (SORD). Earlier studies have confirmed that a hydrogen bonding network in mammalian SORD stabilizes a tetrameric state and is essential for the catalytic power. Our crystal structure does not support such a tetramer formation. We are interested in checking if the oligomer formed in solution matches with our crystal structure.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 项目1 -RNA聚合酶II的转录终止是由一般转录因子IIF（TFIIF）和TFIIF相关的C-末端重复结构域磷酸酶（FCP 1）之间的相互作用促进的。如NMR和CD光谱所示，FCP 1的酸性C-末端区域在游离状态下是无序的，但在与TFIIF的重链结合时采用-螺旋构象。我们最近利用13 C检测的NMR光谱产生全面的NMR化学位移的游离ctFCP 1（残基879-961）和二级位移分析这些数据表明，平衡系综偏向于”，尽管缺乏持久的螺旋结构所示的CD结果。我们的目标是通过在相同的879-961构建体上采集SAXS数据来进一步限制ctFCP 1溶液系综的整体流体动力学形状。 项目2 -我们最近解决了羊肝山梨醇脱氢酶（SORD）的晶体结构。 早期的研究已经证实，哺乳动物SORD中的氢键网络稳定了四聚体状态，并且对于催化能力至关重要。我们的晶体结构不支持这样的四聚体形成。我们感兴趣的是检查在溶液中形成的低聚物是否与我们的晶体结构相匹配。