Co-opting the fermentation pathway for tombusvirus replication: Compartmentalization of cellular metabolic pathways for rapid ATP generation
Fig 8
Confocal microscopy shows co-localization of the co-opted fermention enzymes with the viral repRNAs in whole plants infected with CNV.
(A-B) Most of GFP-AtPdc1 is re-targeted into the replication compartment where the viral RNA synthesis takes place. The viral (+)repRNA carried six copies of the MS2 bacteriophage RNA hairpin (MS2hp), which is recognized by the MS2 coat protein (RFP-MS2-CP). The replication compartment was marked by the BFP-tagged p33 replication protein in N. benthamiana. Panel B shows images from plants mock-inoculated (no viral RNA replication). Note that RFP-MS2-CP contains a week nuclear localization signal, therefore this protein ends up in the nucleus in the absence of replicating (+)repRNA-MS2hp in the cytosol. Expression of the above proteins from the 35S promoter was done after co-agroinfiltration into N. benthamiana leaves. The images were taken 3.5 days after agro-infiltration of plant leaves. Scale bars represent 10 μm. Each experiment was repeated three times. (C-D) Similar experimental set-up as in panel A-B, except the six MS2hps form the suitable structures on the viral (-)repRNA-MS2hp, which is recognized by RFP-MS2-CP. See further details in Panel A. (E-H) Most of GFP-AtAdh1 is re-targeted into the replication compartment where the viral RNA synthesis takes place. See further details in Panel A and C.