Site-specific and mRNA-specific control of accurate mRNA editing by a helicase complex in trypanosomes
| Autor: | Jorge Cruz-Reyes, Andrew Hillhouse, Daniel Camilo Osorio Hurtado, Alasdair Ivens, Xiuren Zhang, Pawan K. Doharey, Joshua Meehan, Zachary T Goodall, Achim Schnaufer, Vikas Kumar, James J. Cai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Trypanosoma
kinetoplastid RNA editing RNA Mitochondrial Protein subunit Trypanosoma brucei brucei Protozoan Proteins Article Substrate Specificity 03 medical and health sciences Ribosomal protein Animals Nucleotide Guide RNA RNA Messenger RNA-Seq Molecular Biology 030304 developmental biology chemistry.chemical_classification 0303 health sciences Messenger RNA biology 030302 biochemistry & molecular biology Helicase RNA Cell biology editosome guide RNA chemistry RNA processing RNA editing biology.protein RNA Editing RNA Protozoan trypanosoma brucei RNA Guide Kinetoplastida REH2C helicase complex |
| Zdroj: | Kumar, V, Ivens, A, Goodall, Z, Meehan, J, Doharey, P K, Hillhouse, A, Hurtado, D O, Cai, J J, Zhang, X, Schnaufer, A & Cruz-Reyes, J 2020, ' Site-specific and mRNA-specific control of accurate mRNA editing by a helicase complex in trypanosomes ', RNA, vol. 26, no. 10, rna.076513.120 . https://doi.org/10.1261/rna.076513.120 RNA |
| Popis: | Trypanosome U-insertion/deletion RNA editing in mitochondrial mRNAs involves guide RNAs (gRNAs) and the auxiliary RNA editing substrate binding complex (RESC) and RNA editing helicase 2 complex (REH2C). RESC and REH2C stably copurify with editing mRNAs but the functional interplay between these complexes remains unclear. Most steady-state mRNAs are partially edited and include misedited “junction” regions that match neither pre-mRNA nor fully edited transcripts. Editing specificity is central to mitochondrial RNA maturation and function, but its basic control mechanisms remain unclear. Here we applied a novel nucleotide-resolution RNA-seq approach to examine ribosomal protein subunit 12 (RPS12) and ATPase subunit 6 (A6) mRNA transcripts. We directly compared transcripts associated with RESC and REH2C to those found in total mitochondrial RNA. RESC-associated transcripts exhibited site-preferential enrichments in total and accurate edits. REH2C loss-of-function induced similar substrate-specific and site-specific editing effects in total and RESC-associated RNA. It decreased total editing primarily at RPS12 5′ positions but increased total editing at examined A6 3′ positions. REH2C loss-of-function caused site-preferential loss of accurate editing in both transcripts. However, changes in total or accurate edits did not necessarily involve common sites. A few 5′ nucleotides of the initiating gRNA (gRNA-1) directed accurate editing in both transcripts. However, in RPS12, two conserved 3′-terminal adenines in gRNA-1 could direct a noncanonical 2U-insertion that causes major pausing in 3′–5′ progression. In A6, a noncanonical sequence element that depends on REH2C in a region normally targeted by the 3′ half of gRNA-1 may hinder early editing progression. Overall, we defined transcript-specific effects of REH2C loss. |
| Databáze: | OpenAIRE |
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