Functional characterization of two paralogs that are novel RNA binding proteins influencing mitochondrial transcripts of Trypanosoma brucei

MRB8170 is incorporated into a macromolecular complex(es) in an RNase-sensitive manner. Mitochondria isolated from 10¹⁰ cells of the T. brucei 29-13 strain were lysed and separated on a 10%–30% glycerol gradient by ultracentrifugation. The gradient was divided into 24 fractions, out of which all odd ones were run on a 10% SDS-acrylamide gel and transferred onto a PVDF membrane that was immunoprobed with the α-MRB8170 antibody. The lysate used in the top panels was treated with RNase inhibitor (RNase-), while that used in the bottom panel was treated with an RNase cocktail that disrupted interactions mediated by both single- and double-stranded RNAs (RNase+).