MOLECULAR RECOGNITION DURING PRE-MRNA SPLICING

Pre-mRNA 剪接过程中的分子识别

• 批准号: 8170296
• 负责人: CLARA KIELKOPF
• 金额: $ 0.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170296
• 关键词: 3' Splice Site; Address; Architecture; Complex; Computer Retrieval of Information on Scientific Projects Database; Funding; Genes; Goals; Grant; Homologous Gene; Housing; Human; Institution; Phosphorylation; Proteins; RNA; RNA Splicing; Recruitment Activity; Research; Research Personnel; Resources; Robotics; SF1; Screening procedure; Signal Transduction; Site; Small Nuclear RNA; Source; Spliceosomes; Staging; Structure; Transcript; United States National Institutes of Health; base; mRNA Precursor; molecular recognition; research study; synchrotron radiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The overall goal of our proposed is to determine the structural basis for 3? splice site recognition. The majority of human gene transcripts are regulated by pre-mRNA splicing. The splice sites are sequentially recognized by protein and RNA components of the spliceosome, a pre-mRNA splicing machine composed of more than 100 proteins and 5 small nuclear RNAs. A complex composed of essential splicing factors SF1, U2AF65, and U2AF35 recognizes the pre-mRNA signals and recruits the core splicing machinery to a target splice site. Specific aims of this proposal address the following central questions concerning the critical early stages of pre-mRNA splicing: (1) What is the structure of a highly-conserved SF1 domain that lacks structural homologues? (2) By what structural means does phosphorylation of this SF1 domain enhance association with U2AF? (3) By what structural means does U2AF adapt to diverse splice sites? (4) What is the three-dimensional architecture of the SF1 / U2AF complex with the target splice site? Synchrotron radiation is essential to address the aims of this proposal for reasons including: (i) Tunable wavelengths are necessary for multiwavelength anomalous dispersion experiments; (ii) The robotic capability greatly facilitates the extensive screening required to identify useful crystals for Aim 1; (iii) Crystal size is small and consequently diffraction is prohibitively weak using conventional in-house x-ray sources.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 我们提议的总体目标是确定3的结构基础？剪接网站识别。大多数人基因转录本受到前MRNA剪接的调节。剪接位点通过蛋白质和剪接体的RNA成分顺序识别，该蛋白质和RNA成分是由100多个蛋白质和5个小核RNA组成的MRNA剪接机。由必需的剪接因子SF1，U2AF65和U2AF35组成的复合物识别前MRNA信号，并将核心剪接机械募集到目标剪接位点。该提案的具体目的解决了以下有关MRNA剪接的关键早期阶段的中心问题：（1）缺乏结构同源物的高度保存的SF1域的结构是什么？ （2）通过该SF1结构域的磷酸化可以增强与U2AF的关联？ （3）U2AF以什么结构含义适应了不同的剪接站点？ （4）SF1 / U2AF复合物具有目标剪接位点的三维体系结构是什么？由于原因包括：（i）可调波长对于多波长异常分散实验是必需的； （ii）机器人能力极大地促进了确定目标1的有用晶体所需的广泛筛选； （iii）晶体尺寸很小，因此使用常规内部X射线源衍射非常弱。