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Fig. S1. Conservation of CosR homologues. Sequence alignment of CosR homologues from related species and characterized MarR-type regulators generated with Culstal W2 (Larkin et al., 2007) and ESPript (Gouet et al., 1999). Residues strictly conserved are indicated with a red background, residues conserved within a group according to a Risler matrix (Risler et al., 1988) are indicated in red and residues conserved between groups are boxed. The following sequences were used to generate the alignment: Vibrio cholerae O1 EI Tor N16961 CosR (NP_230923.1), Vibrio parahaemolyticus (ZP_05119374.1), Vibrio vulnificus (NP_761094.1), Vibrio harveyi (ZP_01988196), Photobacterium profundum (ZP_01222472), Aeromonas hydrophila (YP_858169.1), Methylomicrobium alcaliphilum EctR1 (DQ016501.2), Escherichia coli MarR (NP_416047), Pseudomonas aeruginosa MexR (AAO40258) and Salmonella enterica SlyA (BAA24582.1).

Fig. S2. Growth of ΔcosR, ΔectA and ΔcosRΔectA is similar to wild type. Growth of wild-type, ΔcosR, ΔectA and ΔcosRΔectA strains in (A) 0.2 M NaCl LB or (B) 0.5 M NaCl LB. Strains were diluted 1:200 from overnight cultures into LB containing 0.2 or 0.5 M NaCl and optical density at 600 nm (OD600) measurements were taken at the indicated time points. (C) Growth of wild-type, ΔcosR, ΔectA and ΔcosRΔectA strains in 50% artificial seawater (ASW). Strains were diluted 1:200 from 50% ASW overnight cultures into fresh 50% ASW and serial dilutions of cell culture were plated on LB agar at the indicated time points. Growth is reported as colony forming units (CFU) ml−1. Graphs shown are a representative assay of at least two biological replicates.

Fig. S3. CosR activity is not modulated by ectoine, proline or glycine betaine.

A. β-Galactosidase assays were performed on wild-type, ΔcosR, ΔectA and ΔcosRΔectA strains each containing a chromosomal opuD promoter–lacZ fusion. Strains were grown to exponential phase in LB containing 0.2 and 0.5 M NaCl. The graph shown is a representative assay of three biological replicates and error bars show the standard deviation of eight technical replicates.

B. β-Galactosidase assays were performed on wild-type or ΔcosR strains each containing a chromosomal opuD promoter–lacZ fusion. Cells were grown in LB medium containing 0.2 M NaCl, or 0.2 M NaCl with 25 mM proline (Pro) or 25 mM glycine betaine (GB). The graph shown is a representative assay of three biological replicates and error bars show the standard deviation of eight technical replicates.

Table S1. Bacterial strains and plasmids used in this study.

Table S2. Primers used in this study.

FilenameFormatSizeDescription
emi2805_sm_FS1.eps7120KSupporting info item
emi2805_sm_FS2.eps5132KSupporting info item
emi2805_sm_FS3.eps4098KSupporting info item
emi2805_sm_TS1.doc67KSupporting info item
emi2805_sm_TS2.doc35KSupporting info item

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