Uridine insertion/deletion (U-indel) editing of mitochondrial mRNA, unique to the protistan class Kinetoplastea, generates canonical as well as potentially non-productive editing events. While the molecular machinery and the role of the guide (g) RNAs that provide required information for U-indel editing are well understood, little is known about the forces underlying its apparently error-prone nature. Analysis of a gRNA:mRNA pair allows the dissection of editing events in a given position of a given mitochondrial transcript. A complete gRNA dataset, paired with a fully characterized mRNA population that includes non-canonically edited transcripts, would allow such an analysis to be performed globally across the mitochondrial transcriptome. To achieve this, we have assembled 67 minicircles of the insect parasite Leptomonas pyrrhocoris, with each minicircle typically encoding one gRNA located in one of two similar-sized units of different origin. From this relatively narrow set of annotated gRNAs, we have dissected all identified mitochondrial editing events in L. pyrrhocoris, the strains of which dramatically differ in the abundance of individual minicircle classes. Our results support a model in which a multitude of editing events are driven by a limited set of gRNAs, with individual gRNAs possessing an inherent ability to guide canonical and non-canonical editing.

Department of Biomedical Sciences University of Minnesota Medical School Duluth Campus Duluth MN 55812 USA

Faculty of Biology M 5 Lomonosov Moscow State University Moscow 119991 Russia

Faculty of Science University of South Bohemia 370 05 České Budějovice Czech Republic

Institute for Information Transmission Problems Russian Academy of Sciences Moscow 127051 Russia

Institute of Parasitology Biology Centre Czech Academy of Sciences 370 05 České Budějovice Czech Republic

Life Science Research Centre Faculty of Science University of Ostrava 710 00 Ostrava Czech Republic

Martsinovsky Institute of Medical Parasitology Tropical and Vector Borne Diseases Sechenov University Moscow 119435 Russia

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