RliB interacts with PNPase.

<p>A) Protein affinity purification using 3′ biotinylated RliB. Two biotinylated bait RNAs were used: full length RliB (RliB-B) and a control RNA (<i>S. aureus</i> RNAIII). Coomassie stained SDS-PAGE gel of the two elution fractions is shown. B) Protein affinity purification using 5′ MS2 tagged RliB. Two MS2 tagged bait RNAs were used: full length RliB (RliB-MS2) and a control RNA (<i>S.aureus</i> RsaA). Coomassie stained SDS-PAGE gel of the fractions is shown. C) Interaction between purified PNPase and <i>in vitro</i> transcribed RliB. Gel retardation assay performed with P³²-5′ labelled RliB (RliB) and increasing concentrations of the purified PNPase protein (lane 1, no PNPase added; lane 2, 100 nM PNPase; lane 3, 250 nM PNPase; lane 4, 400 nM PNPase). Competition experiments were performed in the presence of non-labelled RliB (lane 5, 400 nM RliB) and non-labelled RsaA (lane 6, 400 nM RsaA). In these experiments, the concentration of PNPase was 400 nM.