Poster abstracts
Poster number 138 submitted by Rachel Simpson
High Throughput Sequencing of Partially Edited RNAs Provides Insight into the Roles of the TbRGG2 Subcomplex Proteins in Trypanosome RNA editing.
Rachel M. Simpson (Department of Microbiology & Immunology, University at Buffalo School of Medicine), Andrew E. Bruno (Center for Computational Research, University at Buffalo School of Medicine), Jonathan Bard (Next-gen Sequencing Core , University at Buffalo School of Medicine), Runpu Chen, Yijun Sun (Department of Computer Science and Engineering , University at Buffalo School of Medicine), Michael Buck (Department of Biochemistry, University at Buffalo School of Medicine), Laurie K. Read (Department of Microbiology and Immunology, University at Buffalo School of Medicine)
Abstract:
Uridine (U) insertion/deletion RNA editing is an essential process whereby kinetoplastid mitochondrial mRNAs are altered to their final coding forms through sequential utilization of multiple trans-acting gRNAs that act as templates. The enzymes that catalyze U insertion/deletion are part of a multiprotein complex termed the RNA Editing Core Complex (RECC) or editosome. In addition, a large, possibly dynamic and heterogeneous complex termed MRB1 (mitochondrial RNA binding complex 1) is also essential for RNA editing and appears to constitute the platform for editing. Within MRB1 is the incompletely defined TbRGG2 subcomplex, at least some of whose components are essential for the 3’ to 5’ progression of editing. Most RNAs in the steady state population are not completely edited and contain segments of edited RNA between the fully edited and not-yet edited portions of a transcript that match neither and appear to be mis-edited. These segments are termed junction regions, and it is not known whether these regions represent translatable alternative editing, mis-edited sequence that will be corrected, or a terminal error. Using high throughput sequencing, we have analyzed large populations of partially edited sequences for RPS12 and ND7-5’ in parental cells and cells depleted of TbRGG2 subcomplex proteins. We determined that multiple members of the TbRGG2 subcomplex facilitate the progression of editing through a single gRNA and are essential for proper junction formation. Additionally, we found that these proteins have distinct but overlapping roles in facilitating the progression of editing. This study is the first study in which high throughput sequencing is used to elucidate the role of RNA editing factors in Trypanosoma brucei and provides a platform for future research.
Keywords: RNA Editing, trypanosoma, high throughput sequencing