RNA Binding and Core Complexes Constitute the U-Insertion/Deletion Editosome

Author:

Aphasizheva Inna1,Zhang Liye2,Wang Xiaorong3,Kaake Robyn M.3,Huang Lan3,Monti Stefano2,Aphasizhev Ruslan14

Affiliation:

1. Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, Massachusetts, USA

2. Section of Computational Biomedicine, Boston University School of Medicine, Boston, Massachusetts, USA

3. Department of Physiology & Biophysics, School of Medicine, University of California, Irvine, Irvine, California, USA

4. Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts, USA

Abstract

ABSTRACT Enzymes embedded into the RNA editing core complex (RECC) catalyze the U-insertion/deletion editing cascade to generate open reading frames in trypanosomal mitochondrial mRNAs. The sequential reactions of mRNA cleavage, U-addition or removal, and ligation are directed by guide RNAs (gRNAs). We combined proteomic, genetic, and functional studies with sequencing of total and complex-bound RNAs to define a protein particle responsible for the recognition of gRNAs and pre-mRNA substrates, editing intermediates, and products. This approximately 23-polypeptide tripartite assembly, termed the R NA e diting s ubstrate binding c omplex (RESC), also functions as the interface between mRNA editing, polyadenylation, and translation. Furthermore, we found that gRNAs represent only a subset of small mitochondrial RNAs, and yet an inexplicably high fraction of them possess 3′ U-tails, which correlates with gRNA's enrichment in the RESC. Although both gRNAs and mRNAs are associated with the RESC, their metabolic fates are distinct: gRNAs are degraded in an editing-dependent process, whereas edited mRNAs undergo 3′ adenylation/uridylation prior to translation. Our results demonstrate that the well-characterized editing core complex (RECC) and the RNA binding particle defined in this study (RESC) typify enzymatic and substrate binding macromolecular constituents, respectively, of the ∼40S RNA editing holoenzyme, the editosome.

Publisher

American Society for Microbiology

Subject

Cell Biology,Molecular Biology

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