The restoration of surface motility of the rhizobactin-defective mutants was attempted by adding concentrated supernatants containing rhizobactin 1021.
When we examined the number of genes and the GO terms they represent in the double mutants compared to the single mutants, we conclude that Gcn5, Mst2 and Elp3 are functionally redundant in repressing transcription because the number of terms enriched in the double mutants cannot be explained by adding the single mutants' terms together.
While the mutant grew only poorly in CDM (containing 18 mM potassium), normal growth of the trkH mutant was restored by adding potassium at high concentration, establishing the importance of TrkH in potassium acquisition.
This defect in the mutant is countered by adding cobalamin (VitB12) to the medium [169], most likely because the coenzyme B12-dependent ClassII RNRase, NrdJ, assumes the role of NrdDG, while at the same time C. eutrophus H16 readily takes up and assimilates cobalamin.
The D12A/D66A-6His mutant was constructed by adding an oligonucleotide coding for six histidines at the carboxyl end of the enzyme to the plasmid containing the D12A/D66A mutation.
Direct deprotonation may be less likely as we were unable to rescue catalytic activity either with a D495H mutant, or chemically by adding high concentrations of formate or acetate.
Indeed, the observation that the phenotype of disease mutants is somewhat altered by adding the Q67L mutation (Fig. 6A C) provides indirect evidence that a modest amount of GTP hydrolysis does occur in disease mutants in vivo.
Once the selection was finalized, the induction of Flag-TDP-43-12x-Q/N Flag-TDP-43-12x-Q/N Flag-TDP-43-12x-Q/N Flag-TDP-43-12x-Q/N Flag-TDP-43-12x-Q/Nhe culture medium.
However, the growth defects of mitochondrial mutants are not fully reversed by adding exogenous cysteine to the cell cultures: lower cellular densities are reached during the stationary phase than with wild-type cells.
For the insertional mutants, we reconstruct the network by adding the minimal medium input with all energy sources.
