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。这种多样性研究补充建议的总体目标是定义翻译后的作用端粒酶催化蛋白成分，RNA-蛋白质相互作用和端粒酶的修饰布鲁氏锥虫的调节。 T. Brucei是早期发散的寄生虫小ut虫，可以通过人类和昆虫中多种形态上不同的生命周期形成。在T. Brucei中，端粒结构在调节抗原变异方面起着重要作用，使寄生虫能够长期建立感染。端粒酶是T. Brucei中端粒合成的主要调节剂。两个高度保守端粒酶RNA结构结构域，RNA模板和ECR4/5独立结合催化蛋白，端粒合成过程中端粒酶逆转录酶（TERT），是唯一必需的RNA元素用于体外重构催化活性端粒酶。先前关于较高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后翻译后修饰之一端粒酶全酶的成分。但是，泛素化在调节RNA-蛋白质中的作用对于任何真核端粒酶，相互作用和端粒酶活性尚不清楚。最近，我们有表征了t. brucei寄生虫的血液形式的相互作用，作为拟议活动的一部分父母的建议，发现布鲁氏菌端粒酶复合物包含几种泛素化 - 特定的酶和因素。此外，新的实验数据表明，TERT蛋白是差异的泛素化在t. brucei的阶段中形成阶段。因此，在此补充提案，PI打算扩大原始提案的目标，以阐明 tert t. brucei发育过程中TERT和端粒酶RNA的RNA蛋白相互作用。特别是PI计划在TERT蛋白上绘制泛素化位点，并确定其在Brucei T. teromerase功能中的作用。在总而言之，这项研究将奠定PI的长期目标，以定义的核心组成部分端粒长度稳态和基因组完整性的端粒酶激活和相互作用重要的原生物。