RESC14 and RESC8 cooperate to mediate RESC function and dynamics during trypanosome RNA editing.

IF 16.6 2区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Nucleic Acids Research Pub Date : 2024-09-09 DOI:10.1093/nar/gkae561
Katherine Wackowski, Xiaoyu Zhu, Shichen Shen, Ming Zhang, Jun Qu, Laurie K Read
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Abstract

Mitochondrial transcripts in Trypanosoma brucei require extensive uridine insertion/deletion RNA editing to generate translatable open reading frames. The RNA editing substrate binding complex (RESC) serves as the scaffold that coordinates the protein-protein and protein-RNA interactions during editing. RESC broadly contains two modules termed the guide RNA binding complex (GRBC) and the RNA editing mediator complex (REMC), as well as organizer proteins. How the protein and RNA components of RESC dynamically interact to facilitate editing is not well understood. Here, we examine the roles of organizer proteins, RESC8 and RESC14, in facilitating RESC dynamics. High-throughput sequencing of editing intermediates reveals an overlapping RESC8 and RESC14 function during editing progression across multiple transcripts. Blue native PAGE analysis demonstrates that RESC14 is essential for incorporation of RESC8 into a large RNA-containing complex, while RESC8 is important in recruiting a smaller ribonucleoprotein complex (RNP) to this large complex. Proximity labeling shows that RESC14 is important for stable RESC protein-protein interactions, as well as RESC-RECC associations. Together, our data support a model in which RESC14 is necessary for assembly of editing competent RESC through recruitment of an RNP containing RESC8, GRBC and gRNA to REMC and mRNA.

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在锥虫RNA编辑过程中,RESC14和RESC8合作介导RESC的功能和动态。
布氏锥虫的线粒体转录本需要大量尿苷插入/缺失 RNA 编辑才能生成可翻译的开放阅读框。RNA 编辑底物结合复合物(RESC)是在编辑过程中协调蛋白质-蛋白质和蛋白质-RNA 相互作用的支架。RESC 大致包含两个模块,即引导 RNA 结合复合物(GRBC)和 RNA 编辑中介复合物(REMC),以及组织者蛋白。目前还不太清楚 RESC 的蛋白质和 RNA 成分如何动态地相互作用以促进编辑。在这里,我们研究了组织者蛋白RESC8和RESC14在促进RESC动态过程中的作用。编辑中间产物的高通量测序揭示了 RESC8 和 RESC14 在多个转录本的编辑过程中的重叠功能。蓝色原生 PAGE 分析表明,RESC14 是将 RESC8 并入大型含 RNA 复合物的必要条件,而 RESC8 则是将较小的核糖核蛋白复合物(RNP)招募到该大型复合物的重要条件。接近标记显示,RESC14 对稳定的 RESC 蛋白-蛋白相互作用以及 RESC-RECC 关联非常重要。总之,我们的数据支持这样一个模型:RESC14 是通过将包含 RESC8、GRBC 和 gRNA 的 RNP 招募到 REMC 和 mRNA 上,从而组装出具有编辑能力的 RESC 的必要条件。
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来源期刊
Nucleic Acids Research
Nucleic Acids Research 生物-生化与分子生物学
CiteScore
27.10
自引率
4.70%
发文量
1057
审稿时长
2 months
期刊介绍: Nucleic Acids Research (NAR) is a scientific journal that publishes research on various aspects of nucleic acids and proteins involved in nucleic acid metabolism and interactions. It covers areas such as chemistry and synthetic biology, computational biology, gene regulation, chromatin and epigenetics, genome integrity, repair and replication, genomics, molecular biology, nucleic acid enzymes, RNA, and structural biology. The journal also includes a Survey and Summary section for brief reviews. Additionally, each year, the first issue is dedicated to biological databases, and an issue in July focuses on web-based software resources for the biological community. Nucleic Acids Research is indexed by several services including Abstracts on Hygiene and Communicable Diseases, Animal Breeding Abstracts, Agricultural Engineering Abstracts, Agbiotech News and Information, BIOSIS Previews, CAB Abstracts, and EMBASE.
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