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Administrative Supplement to Promote Diversity in DecodingRNA-Protein Interactions in Trypanosoma Telomerase

促进锥虫端粒酶 RNA-蛋白质相互作用解码多样性的行政补充

基本信息

批准号：
10797998
负责人：
Kausik Chakrabarti
金额：
$ 13.1万
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：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2025-04-30
项目状态：
未结题

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. The overall goal of this diversity research supplement proposal is to define the role of post-translational modifications of the telomerase catalytic protein component, TERT in RNA-protein interactions and telomerase regulation in Trypanosoma brucei. T. brucei is 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. Telomerase is the major regulator 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. Previous studies on higher eukaryotes revealed that ubiquitination is one of the major post-translational modifications that affect the stability and activity of TERT component in telomerase holoenzyme. However, the role of ubiquitination in regulating RNA-protein interactions and telomerase activity remains unknown for any eukaryotic telomerase. Recently, we have characterized the interactome of the bloodstream form T. brucei parasites as part of the proposed activities in the parent proposal and found that T. brucei telomerase enzyme complex contains several ubiquitination - specific enzymes and factors. Additionally, new experimental data revealed that TERT protein is differentially ubiquitinated in the procyclic and bloodstream form developmental stages of T. brucei. Therefore, in this supplement proposal, PI intends to expand the original proposal’s objectives to elucidate the mechanisms of RNA-protein interactions of TERT and telomerase RNA during T.brucei development. Particularly, PI plans to map the ubiquitination sites on the TERT protein and determine their roles in T. brucei telomerase function. 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，它可以从端粒复制端粒重复序列，在真核生物中，RNA是一个完整的RNA组分，它可以被连接到线性染色体的末端，以防止基因组的不稳定性。的本多样性研究补充提案的总体目标是定义翻译后的作用 RNA-蛋白质相互作用中的端粒酶催化蛋白组分、端粒酶的修饰和端粒酶布氏锥虫中的调节。T.布氏杆菌是一种早期的分歧寄生原生生物，在人类和昆虫中有多种形态上不同的生命周期形式。于T.布氏杆菌的端粒结构在抗原变异的调节中起着重要作用，使寄生虫能够建立长期的感染端粒酶是T.布鲁塞。两个高度保守端粒酶RNA结构域、RNA模板和eCR 4/5独立地结合催化蛋白，端粒酶逆转录酶（TERT）是端粒合成过程中唯一必需的RNA元件用于催化活性端粒酶的体外重建。先前对高等真核生物的研究表明，泛素化是影响TERT稳定性和活性的主要翻译后修饰之一端粒酶全酶的组成部分。然而，泛素化在调节RNA蛋白中的作用，对于任何真核端粒酶，相互作用和端粒酶活性仍然未知。最近我们其特征在于血液形式T的相互作用组。关于将布鲁氏菌寄生虫病作为父母的建议，并发现T。布氏杆菌端粒酶复合物含有几个泛素化位点，特定的酶和因子。此外，新的实验数据显示，TERT蛋白质在细胞内的表达存在差异。在T.布鲁塞。所以在这补充建议，PI打算扩大原来的建议的目标，阐明机制，布氏锥虫发育过程中端粒酶和端粒酶RNA的相互作用。特别是，PI计划绘制TERT蛋白上的泛素化位点，并确定它们在T.布氏杆菌端粒酶功能在总之，这项研究将为PI的长期目标奠定基础，以确定核心组成部分，端粒酶激活与端粒长度稳态和基因组完整性相互作用重要的原生生物