Co-opting the fermentation pathway for tombusvirus replication: Compartmentalization of cellular metabolic pathways for rapid ATP generation
Fig 11
The co-opted cytosolic Pdc1 fermentation enzyme affects ATP accumulation within the tombusvirus replication compartment in yeast.
(A) A scheme of the FRET-based detection of ATP within the tombusvirus replication compartment. The enhanced ATP biosensor, ATeamYEMK was fused to TBSV p92pol replication protein. See further details in the main text. (B) Comparison of the ATP level produced within the tombusvirus replication compartment in pdc1Δ yeast strain expressing WT Pdc1p, Pdc1S455F mutant or without Pdc1 expression using ATeamYEMK -p92pol. The more intense FRET signals are white and red (between 0.5 to 1.0 ratio), whereas the low FRET signals (0.1 and below) are light blue and dark blue. We show the quantitative FRET values (obtained with ImageJ) for a number of samples in the graph. Note that we also used a reduced ATP-sensitive version of ATeamRK-p92 (bottom panel) to demonstrate that the FRET signal is due to ATP-sensing. Scale bars represent 5 μm. Each experiment was repeated three-four times.