Table S1. Oligonucleotides used as PCR primers.


Fig. S1. Cell growth in BSM succinate 5 μM. Wild-type PAO1 cells were grown in BSM succinate 5 μM (black diamonds) and in water as negative control for contaminations (white diamonds). Cell growth was monitored by plating an opportune dilution of the sample at different time points and by counting the number of colony-forming units (cfu). The initial inoculum was 50 cells ml−1. Each value is the average of three different cultures ± standard deviation. For every time points triple technical replicates were performed. In some instances, the standard deviation bars are smaller than the symbols used.


Fig. S2. Effect of succinate concentrations on the levels of CrcZ. Northern blots of PAO1 wild type grown in BSM succinate 5 μM (lane 1) and 40 mM (lane 2) for about 3 h. Three micrograms of total RNA was loaded onto a 12% polyacrylamide gel. CrcZ and the 5S rRNA were detected using a digoxigenin (DIG)-labelled double-stranded probe. The signal of 5S rRNA was used as a loading control.


Fig. S3. Transcriptional regulation of crcZ expression by Crc. Expression of a crcZ–gfp reporter fusion (pME9824) was measured in PAO1 wild type (black bars) and PAO1 Δcrc mutant PAO6673 (white bars) grown to OD600 ≈ 1.5 in BSM mannitol, glucose or succinate 40 mM. Relative fluorescence was corrected for background as described in Experimental procedures. Each value is the average of three different cultures ± standard deviation.

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