title: Investigating RNA editing factors from trypanosome mitochondria.
abstract: Mitochondrial U-insertion/deletion mRNA editing is carried out by two principal multiprotein assemblies, enzymatic RNA editing core (RECC) and RNA editing substrate binding (RESC) complexes, and a plethora of auxiliary factors. An integral part of mitochondrial gene expression, editing receives inputs from primary mRNA and gRNA precursor processing pathways, and generates substrates for mRNA polyadenylation and translation. Although nearly all RECC-embedded enzymes have been implicated in specific editing reactions, the majority of proteins that populate the RESC are also essential for generating edited mRNAs. However, lack of recognizable motifs in RESC subunits limits the prowess of bioinformatics in guiding biochemical experiments and elucidating their specific biological functions. In this chapter, we describe a generic workflow for investigating mitochondrial mRNA editing in Trypanosoma brucei and focus on several methods that proved instrumental is assigning definitive functions to editing factors lacking known signature sequences.
authors.affiliation: Department of Molecular and Cell Biology, Boston University School of Dental Medicine, Boston, MA 02118, USA. Electronic address: innaaf@bu.edu.
authors.forename: Inna
authors.initials: I
authors.lastname: Aphasizheva
authors.affiliation: Section of Computational Biomedicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Liye
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Molecular and Cell Biology, Boston University School of Dental Medicine, Boston, MA 02118, USA; Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Ruslan
authors.initials: R
authors.lastname: Aphasizhev
mesh_terms: D000818:Animals; D008928:Mitochondria / Q000235:genetics*; D008967:Molecular Biology / Q000379:methods*; D026723:Polyadenylation / Q000235:genetics; D017393:RNA Editing / Q000235:genetics*; D017394:RNA, Guide, Kinetoplastida / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D014346:Trypanosoma brucei brucei / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D017394:RNA, Guide, Kinetoplastida
keywords: Guide RNA; Mitochondria; Protein complexes; RNA editing; RNA ligase; TUTase; Trypanosoma