Guide RNA Binding Complex

引导RNA结合复合物

• 批准号: 9352420
• 负责人: Ruslan Afasizhev
• 金额: $ 51.56万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-10 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9352420
• 关键词: Antigenic Variation; Antisense RNA; Architecture; Area; Binding; Binding Sites; Biology; Characteristics; Chemicals; Complex; Cryoelectron Microscopy; Crystallization; DNA; DNA-Directed RNA Polymerase; Data; Development; Diphosphates; Ensure; Event; Excision; Exonuclease; Funding; Gap Junctions; Genes; Genetic; Genetic Transcription; Genome; Goals; Guide RNA; Health Hazards; Holoenzymes; Hybrids; Hydrolysis; Individual; Investigation; Kinetoplast DNA; Knowledge; Maps; Mass Spectrum Analysis; Mediator of activation protein; Medical; Messenger RNA; Methods; Mitochondria; Mitochondrial RNA; Modality; Modeling; Modification; Molecular; Parasites; Pathway interactions; Pattern; Polyadenylation; Polynucleotide Adenylyltransferase; Population; Positioning Attribute; Pre-mRNA Polyadenylation Factor; Process; Property; Proteins; RNA; RNA Binding; RNA Editing; RNA Processing; Reaction; Recruitment Activity; Research; Resolution; Ribosomes; Role; Specificity; Structure; System; Tail; Testing; Transcript; Transcription Initiation Site; Translational Activation; Translations; Tropical Disease; Trypanosoma; Trypanosoma brucei brucei; base; crosslink; endonuclease; experimental study; genome-wide analysis; health economics; in vivo; insertion/deletion mutation; mRNA Precursor; mRNA Stability; multimodality; new therapeutic target; novel; pathogen; polypeptide; programs; promoter; reconstitution; sensor; transcriptome; tripolyphosphate

ABSTRACT Trypanosoma brucei species inflict health hazards and economic hardship on arguably the most marginalized populations in the world. Some of the best-studied Excavata, trypanosomes also represent important models in many areas of research, including antigenic variation, host-pathogen interaction, developmental reprogramming and mitochondrial biology. This project will elucidate mechanisms by which macromolecular RNA editing substrate binding complex (RESC) stabilizes and delivers mitochondrial pre-mRNAs and guide RNAs into the U-insertion/deletion editing pathway, and coordinates polyadenylation and translation of edited mRNAs. We establish the RESC platform as the RNA binding constituent of the editing holoenzyme and seek to investigate its role in editing reactions, and functions beyond the RNA editing process. To this end, we demonstrate that RESC-associated MERS1 pyrophosphohydrolase and KPAP1 poly(A) polymerase target pre-mRNA 5′ and 3′ ends, respectively. Importantly, both 5′ pyrophosphate removal and 3′ A-tailing appear to be critical for pre-mRNA stabilization prior to editing. Conversely, specific module within RESC is suggested to couple the completion of editing with post-editing 3′ A/U-tailing and mRNA binding to the ribosome. Collectively, the existing evidence positions the ~25 polypeptide RESC complex as the multimodal nexus of mitochondrial RNA processing. Furthermore, initial investigation of RESC-associated MERS1 complex, RNA polymerase (MTRNAP), and the 3′ processome (MPsome) challenges the long-standing model of multicistronic maxicircle transcription and endonucleolytic partitioning of primary transcripts. The proposed experiments will deepen understanding of RNA editing by determining the RESC structure at near-atomic resolution and RNA binding specificities of individual subunits. We will test a broad functional hypothesis that discrete RESC modules coordinate completion of mRNA editing with 3′ modification and translational activation. Finally, we put forward a fundamentally novel concept of monocistronic pre-mRNAs that are transcribed from individual promoters and shaped by 5ʹ modification and antisense RNA-controlled 3′-5′ degradation. By elucidating the RESC structure, RNA binding properties, and higher-order interactions, and evaluating the paradigm-shifting “monocistronic hypothesis,” this program will expand the knowledge of critical parasite-specific processes and may provide new drug targets.

摘要