FIG 5.

The metabolite methylmalonic acid is an inhibitor of PccD. (A) EMSA of His-PccD protein binding to the upstream promoter regions of pccBC. The biotin-labeled DNA probe (about 15 ng, reaction system 10 μl) was incubated with 5 μM His-PccD. Propionate, propionyl-CoA, methylmalonic acid, methylmalonyl-CoA, or succinyl-CoA was added as a cofactor to a final concentration of 1.0 mM. (B) Total RNA of S. erythraea NRRL2338 was extracted after 36 h of growth in TSB medium or TSB supplied with 10 mM methylmalonic acid. Relative transcript levels were normalized to the 16S rRNA. The transcription value of pccBC in S. erythraea WT cultivated in TSB medium was arbitrarily normalized to 1. Error bars represent the standard deviations from three biological replicates.