Trypanosomatid, fluorescence-based in vitro U-insertion/U-deletion RNA-editing (FIDE).

ABSTRACT: Gene expression within the mitochondria of African trypanosomes and other protozoan organisms relies on a nucleotide-specific RNA-editing reaction. In the process exclusively uridine (U)-nucleotides are site-specifically inserted into and deleted from sequence-deficient primary transcripts to convert them into translatable mRNAs. The reaction is catalyzed by a 0.8 MDa multiprotein complex termed the editosome. Here we describe an improved in vitro test to quantitatively explore the catalytic activity of the editosome. The assay uses synthetic, fluorophore-derivatized oligoribonucleotides as editing substrates, which enable the automated electrophoretic separation of the reaction products by capillary electrophoresis (CE) coupled to laser-induced fluorescence (LIF) detection systems.

SUBMITTER: Leeder WM 

PROVIDER: S-EPMC8008098 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature