Fig. 3: Single-molecule simultaneous tracking of target RNA and Vasa dissociation.
From: Heterogeneous dissociation process of truncated RNAs by oligomerized Vasa helicase
a The fluorescent images of target RNA (magenta) colocalized with the oligomerized WT-Vasa or dN-Vasa (green) in the presence of Siwi. Scale bars represent 10 µm. b The distribution of the number of TMR-Vasa molecules associated with a single molecule of target RNA, calculated from the fluorescence intensity (N = 276). The molecular numbers of Vasa and RNA were calculated from the averaged intensity of the respective single fluorescent dye (b–e, g). Inset shows the schematic representation of the complex with target RNA and Vasa loaded on Siwi-piRISC. c–e Time-trace intensities of the colocalized Vasa (upper, green) and cleaved RNA (lower, red) dissociating after adding 1 mM ATP. The intensity step was analysed, and the change in the molecular number of each factor converted from the fluorescence intensity was illustrated. c Vasa and RNA dissociated simultaneously. d Vasa partially dissociated independently of RNA dissociation. e Vasa partially dissociated but RNA kept binding. f Classify the dissociation of Vasa and RNA according to (c–e). The counts of Vasa dissociation completely are given in brackets. g Vasa dissociated partially as a single molecule (85%) in the colocalized spots (96 data in 52 spots).
