Decoding RNA-Protein Interactions in Trypanosoma Telomerase

解码锥虫端粒酶中 RNA-蛋白质相互作用

基本信息

批准号：
10515146
负责人：
Kausik Chakrabarti
金额：
$ 47.44万
依托单位：
UNIVERSITY OF NORTH CAROLINA CHARLOTTE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10515146
关键词：
Active Sites Adopted Affect Antigenic Variation Binding Biochemical Biology Cell Proliferation Cell Shape Cells Chromosomes Clinical Complex DNA Data Detection Development Disease Elements Enzymes Eukaryota Foundations Gene Expression Genes Genomic Instability Goals Heterogeneous-Nuclear Ribonucleoprotein Group F-H Heterogeneous-Nuclear Ribonucleoproteins Holoenzymes Homeostasis Human Immune In Vitro Infection Insecta Length Life Life Cycle Stages Mass Spectrum Analysis Mediating Modeling Modification Molecular Molecular Chaperones Molecular Conformation Morphology Multienzyme Complexes Mutation Neurologic Nuclear Nuclear Proteins Nucleotides Parasite Control Parasites Pathogenesis Pathway interactions Play Proliferating Protein Family Proteins Proteomics RNA RNA Folding RNA Stability RNA-Directed DNA Polymerase RNA-Protein Interaction Regulation Research Ribonucleoproteins Role Site Structure Telomerase Telomerase RNA Component Telomere Maintenance Trypanosoma Trypanosoma brucei brucei Tumor stage Virulence Yeasts base genome integrity novel pathogen prevent reconstitution response scaffold telomere

项目摘要

Project Summary Telomerase is a unique ribonucleoprotein enzyme that processively adds telomeric repeats, copied from its integral RNA component, to the ends of linear chromosomes to prevent genome instability in eukaryotes. This proposal seeks to define RNA folding and RNA-protein interactions that are critically important for telomerase regulation in Trypanosoma brucei, an early divergent parasitic protist that proliferates through multiple morphologically distinct life cycle forms in humans and insects. In T. brucei, the telomere structure plays an important role in regulation of antigenic variation that enables the parasite to establish a long-term infection. Particularly, extremely short telomeres could jeopardize telomere integrity, stability of their (sub)telomeric virulence genes and parasite survival. Therefore, understanding the mechanism that controls telomere replication in T. brucei could provide important clues to control parasite proliferation. Telomerase is the major mechanism of telomere synthesis in T. brucei. Two highly conserved telomerase RNA structural domains, the RNA template and eCR4/5 independently bind the catalytic protein, telomerase reverse transcriptase (TERT) during telomere synthesis and are the only required RNA elements for in vitro reconstitution of catalytically active telomerase. However, T.brucei telomerase RNA has unusual sequence and structural composition in the above domains compared to ciliate, yeast and vertebrate telomerase RNAs, suggesting novel modes of regulation for telomere synthesis. Therefore, our hypothesis is that these unusual sequence and structural diversity of T. brucei telomerase RNA domains cause differences in RNA-protein interactions and conformational changes, resulting in species-specific telomerase assembly and activity. Our recent RNA structure probing data from two replicative stages of T. brucei suggests that RNA folding and telomerase activation could be developmentally regulated. To understand how stage -specific structural rearrangements and RNA-proteins interactions control telomerase regulation in T. brucei, in Aim 1 of the proposal we will determine molecular requirements of RNA-protein interactions in the above two domains in T. brucei telomerase. Aim 2 of this proposal will explore additional requirements for telomerase regulation by dissecting RNA-specific factors that are required for functional telomerase RNP assembly and activity. In summary, this research will lay the foundation for the PI's long-term goal to define core components of telomerase activation and interactions for telomere length homeostasis and genome integrity in a clinically important protist.

项目摘要端粒酶是一种独特的核糖核蛋白酶积分RNA成分，到线性染色体的末端，以防止真核生物中的基因组不稳定性。这提案旨在定义对端粒酶至关重要的RNA折叠和RNA - 蛋白质相互作用 Brucei锥虫瘤的调节，这是一种早期发散的寄生虫原生物，通过多个增殖人类和昆虫中形态上不同的生命周期形式。在T. Brucei中，端粒结构播放在调节抗原变异方面的重要作用，使寄生虫能够建立长期感染。特别是，极短的端粒可能会危害端粒完整性，端粒的稳定性毒力基因和寄生虫存活。因此，了解控制端粒的机制布鲁氏菌中的复制可以提供重要的线索来控制寄生虫增殖。端粒酶是主要的 T. Brucei中端粒合成的机理。两个高度保守的端粒酶RNA结构结构域， RNA模板和ECR4/5独立结合催化蛋白，端粒酶逆转录酶（TERT）在端粒合成期间，这是催化性重构的唯一必需的RNA元素活性端粒酶。然而，乳杆菌端粒酶RNA在与纤毛，酵母和脊椎动物端粒酶RNA相比调节端粒合成。因此，我们的假设是这些不寻常的序列和结构 t. brucei端粒酶RNA结构域的多样性导致RNA-蛋白质相互作用和构象变化，导致物种特异性的端粒酶组装和活性。我们最近的RNA 从T. brucei的两个复制阶段探测数据的结构表明RNA折叠和端粒酶激活可以受到发展调节。了解阶段特定的结构重排如何 RNA - 蛋白质相互作用控制T. Brucei中的端粒酶调节，在该提案的AIM 1中，我们将确定上述两个域中RNA蛋白相互作用的分子需求端粒酶。该提案的目标2将通过解剖探索端粒酶调节的其他要求功能端粒酶RNP组装和活性所需的RNA特异性因子。总而言之，这研究将奠定PI的长期目标，以定义端粒酶激活的核心组成部分以及端粒长度稳态和基因组完整性的相互作用。