Fig. 5: CPAC components are depleted from cleavage bodies and redistribute to the RNAPII compartment in the absence of Dbp2. | Nature Communications

Fig. 5: CPAC components are depleted from cleavage bodies and redistribute to the RNAPII compartment in the absence of Dbp2.

From: DEAD-box ATPase Dbp2 is the key enzyme in an mRNP assembly checkpoint at the 3’-end of genes and involved in the recycling of cleavage factors

Fig. 5

A Live cell imaging for Pcf11-GFP or Rna15-GFP in wild type (marked by NLS-BFP) mixed with P.nmt-dbp2. Cells were shifted to YES for 6 h. The GFP channel is shown as the maximal intensity projection using a blue orange icb colour scale as indicated. Mixed wild type (+/- BFP) was included as control. Extended fields of view are provided in Suppl. Fig. 5A, B. B Quantification of the experiment shown in (A) (n = 2 independent experiments). Measurements were performed on maximal intensity Z-projections and the maximal green fluorescence intensity scored for each cell. Mixed cultures of wild types were included as controls. Numbers below the graph indicate number of counted cells for each condition. Box plot features as in Fig. 3E. The displayed p-values for pair-wise comparisons were calculated using a two-sided Wilcoxon rank sum test. Source data are provided as a Source Data file. C Live cell imaging of genomically tagged Pcf11-GFP and the MTREC component Red1-tdTomato. Cells were shifted to YES for 6 h. The merged channel shows Pcf11-GFP in green and Red1-tdTomato in magenta. Images are representative of three independent experiments. Fluorescence intensity profiles along the yellow lines are shown in (D). D Fluorescence intensity profiles as indicated in (C). Green line corresponds to the Pcf11-GFP signal, dashed magenta line to the Red1-tdTomato signal. Fluorescence intensities were normalised to a 0–1 range. Source data are provided as a Source Data file. E Live cell imaging of genomically tagged Pcf11-GFP and the RNAPII component Rpb9-iRFP. Cells were shifted to YES for 6 h. The merged channel shows Pcf11-GFP in green and Rpb9-iRFP in magenta. Images are representative of two independent experiments. Fluorescence intensity profiles along the yellow lines are shown in F. F Fluorescence intensity profiles as indicated in (E). Green line corresponds to the Pcf11-GFP signal, dashed magenta line to the Rpb9-iRFP signal. Source data are provided as a Source Data file.

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