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emi412123-sup-0001-figs1.eps370KFig. S1. Comparison of activity between the BACTH pKT25 and pST25 constructs. Test of interactions between E. coli FtsW (W) and FtsI (I) using the T25 vector pKT25 or pST25 versus pUT18C. The labels are in the format T25/T18 (e.g. W/I = T25-FtsW versus T18-FtsI). Neg = negative control consisting of pooled data from tests of constructs versus empty vectors or unrelated chlamydial inner membrane proteins, Ct471 or GlnP. β-galactosidase activity was measured from eight colonies from each of two experiments and is expressed as average relative units with standard deviation. A positive interaction is considered if the test interaction has fivefold greater activity compared with negative controls.
emi412123-sup-0002-figs2.eps390KFig. S2. Validation of BACTHGW system using eukaryotic and viral protein–protein interactions. The labels are in the format T25/T18 (e.g. 16E6/IRF3 = T25-16E6 versus T18-IRF3). 16E6 = human papillomavirus 16 E6 protein. IRF3 = interferon regulatory factor 3. Smad2, Smad3 and Smad4 are human signal transduction proteins requiring post-translation modification to interact. Pos = E. coli T25-FtsW versus T18-FtsI. Neg = pooled data from tests of constructs versus empty vectors or unrelated chlamydial inner membrane proteins, Ct471 or GlnP/Ct129. The experiments were performed as described in the figure legend for Fig. S1.
emi412123-sup-0003-figs3.eps2497KFig. S3. topcons transmembrane domain analysis of the hypothetical chlamydial proteins Ct471 and Ct605. Note the relatively low confidence score of Ct605 compared with Ct471 and the corresponding data in Fig. 3 of the text.
emi412123-sup-0004-tables1.docx116KTable S1. List of primers and plasmids used.

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