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The following supplementary material is available for this article: Table S1. Pimers used in this study. Download here. Fig. S1. Growth phenotype of P. putida DS1 mutants. P. putida strains DS1 (wild-type), Dfi74J (sfnR::Tn5), SRK1 (sfnR::kan), and SRK1DMSO2+ (sfnR::kan, mini-Tn5) were grown on DMSO2 as the sole sulphur source. Fig. S2. Complementation of SRK1 with a plasmid producing Sfn proteins. pBBad22T (vector control), pEN20 (producing SfnF and SfnG), pEN19 (producing SfnA and SfnB), pEN04 (producing SfnA), and pEN10 (producing SfnB) were respectively introduced into SRK1. The transformants were grown on DMSO2 as a sulphur source. Download here. Fig. S3. Primer extension analysis. The total RNA from P. putida DS1 grown exponentially (OD600< 0.95) on sulphate or DMSO2 was subjected to a reverse transcriptional reaction with sfnF-PE2-RV primer (Table S1). Lane 1, total RNA from P. putida DS1 cells grown on sulfate; lane 2, total RNA from cells grown on DMSO2. The sequence ladder obtained with the primer sfnF-PE2-RV is shown on the left. Fig. S4. Gel mobility shift assay. A. Physical map of the sfnA-sfnF intergenic region and probes (bidirectional arrows). The SfnR binding sites shown in the result of DNase I footprinting are shown below the physical map. The closed box on the physical map indicates the 7-bp-long sequence (5'-GCTGTTG-3'). B. Gel mobility shift assay using purified N-ht-SfnR. Pb3 and Pb6, containing the 7-bp-long sequence, showed no retarded bands, indicating that the 7-bp-long sequence itself is not required for the binding of SfnR. Pb4 and Pb5, both of which contained the 7-bp-long sequence, showed a retarded signal, suggesting that the site 2 and site 3 regions are not overlapped and that the flanking sequence of the 7-bp-long sequence is required for the binding of SfnR. Download here

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