Regulatory functions of intrinsically disordered electronegative clusters (ENC) in RNA-binding proteins

RNA结合蛋白中本质无序的负电簇（ENC）的调节功能

基本信息

批准号：
2024964
负责人：
Jun Zhang
金额：
$ 90万
依托单位：
University of Alabama at Birmingham
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2024964&HistoricalAwards=false
关键词：
Regulatory functions intrinsically disordered electronegative

项目摘要

Interaction between biomolecules is essential for life. While most proteins have stable three-dimensional structures, a significant number of proteins have been shown to lack a stable three-dimensional structure and are known as intrinsically disordered proteins. It has been shown that proteins that interact with RNA contain regions that are intrinsically disordered, which are essential for the protein-RNA binding. The investigators of this project have shown that most abundant disordered regions in RNA-binding proteins encode clusters of acidic residues or phosphorylation sites that give these regions an overall negative charge. Like tails that help animals to balance and to swish away insects, the disordered protein regions help proteins to balance surface charges for stability and swish away nonspecific ligands. This project will explore how these electronegative clusters modulate protein function. Studying these proteins will provide insights into key biological processes, such as transient folding of disordered proteins, site selection in ribosomal biogenesis, and alternative splicing. In addition, understanding this phenomenon will inform development of novel protein design techniques for more robust and specific RNA-binding proteins, which will ultimately contribute to US bioeconomy. This project will provide training opportunities to high school and undergraduate students, including members of underrepresented groups, in biochemical research and bioinformatics-based coding. This project will study three model proteins: histone-mRNA Stem-Loop Binding Protein, ribosome biogenesis protein 15, and Serine-Arginine Rich Splicing Factor 1. These proteins were selected because they play essential roles in critical biological processes, such as histone pre-mRNA processing, ribosomal biogenesis, and alternative splicing, respectively. Because these proteins encode different types of electronegative clusters and have unique degrees of three-dimensional folding status, this project will systematically elucidate the function of electronegative clusters in mediating both protein folding and stability as well as RNA binding specificity and kinetics. Cutting-edge biophysical and biochemical techniques, such nuclear magnetic resonance, crystallography, stopped-flow fluorescence, and next generation sequencing will be used. Completion of this work will not only produce knowledge generally applicable to all RNA-binding proteins, but also elucidate specific and important processes where these proteins play essential roles.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

生物分子之间的相互作用对于生命至关重要。虽然大多数蛋白质具有稳定的三维结构，但大量蛋白质已被证明缺乏稳定的三维结构，被称为本质上无序的蛋白质。研究表明，与 RNA 相互作用的蛋白质含有本质上无序的区域，这对于蛋白质与 RNA 的结合至关重要。该项目的研究人员表明，RNA 结合蛋白中最丰富的无序区域编码酸性残基簇或磷酸化位点，使这些区域总体带有负电荷。就像帮助动物保持平衡和驱走昆虫的尾巴一样，无序的蛋白质区域帮助蛋白质平衡表面电荷以保持稳定性并驱走非特异性配体。该项目将探索这些负电簇如何调节蛋白质功能。研究这些蛋白质将有助于深入了解关键的生物过程，例如无序蛋白质的瞬时折叠、核糖体生物发生中的位点选择以及选择性剪接。此外，了解这一现象将为开发新的蛋白质设计技术提供信息，以开发更强大和更特异的 RNA 结合蛋白，这最终将为美国生物经济做出贡献。该项目将为高中生和本科生（包括代表性不足群体的成员）提供生化研究和基于生物信息学的编码方面的培训机会。该项目将研究三种模型蛋白：组蛋白-mRNA 干环结合蛋白、核糖体生物发生蛋白 15 和富含丝氨酸-精氨酸的剪接因子 1。选择这些蛋白是因为它们在关键的生物过程中发挥重要作用，例如组蛋白预分别是 mRNA 加工、核糖体生物合成和选择性剪接。由于这些蛋白质编码不同类型的负电簇，并具有独特程度的三维折叠状态，本项目将系统地阐明负电簇在介导蛋白质折叠和稳定性以及RNA结合特异性和动力学中的功能。将使用尖端的生物物理和生化技术，如核磁共振、晶体学、停流荧光和下一代测序。这项工作的完成不仅将产生普遍适用于所有 RNA 结合蛋白的知识，而且还将阐明这些蛋白质发挥重要作用的特定和重要过程。该奖项反映了 NSF 的法定使命，并通过使用基金会的评估进行评估，认为值得支持。智力价值和更广泛的影响审查标准。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Inter-domain Flexibility of Human Ser/Arg-Rich Splicing Factor 1 Allows Variable Spacer Length in Cognate RNA’s Bipartite Motifs

人类富含 Ser/Arg 的剪接因子 1 的域间灵活性允许同源 RNA 的二分基序中的间隔区长度可变

DOI：
10.1021/acs.biochem.2c00565
发表时间：
2022-12
期刊：
Biochemistry
影响因子：
2.9
作者：
De Silva, Naiduwadura Ivon;Fargason, Talia;Zhang, Zihan;Wang, Ting;Zhang, Jun
通讯作者：
Zhang, Jun

Electronegative clusters modulate folding status and RNA binding of unstructured RNA‐binding proteins

负电簇调节非结构化 RNA 结合蛋白的折叠状态和 RNA 结合