Fig. 3: Tme/i1 are an antibacterial BB22OP T6SS1 E/I pair.
From: A comparative genomics methodology reveals a widespread family of membrane-disrupting T6SS effectors
a Gene structure of tme/i1. Genes are represented by arrows indicating the direction of translation. White arrows denote genes unrelated to T6SS. Locus tags (VPBB_RSxxxxx) are shown above. b Expression (cells) and secretion (media) of C-terminally Flag-tagged Tme1 from T6SS1+ (Δhns) and T6SS1− (Δhns/Δhcp1) V. parahaemolyticus BB22OP strains were detected by immunoblotting using anti-Flag antibodies. Tme1-Flag was expressed from an arabinose-inducible plasmid (pTme1). Loading control (LC), visualized as Ponceau S stained membrane, is shown for total protein lysates. The experiment was independently repeated three times with similar results. Results from a representative experiment are shown. c, d Viability counts of V. parahaemolyticus BB22OP prey before (0 h) and after (4 h) co-incubation with the indicated V. parahaemolyticus BB22OP attackers on media containing L-arabinose at 30 °C (n = 3 co-cultures). Δhns was used as the parental T6SS1+ strain, and Δhns/Δhcp1 was used as a T6SS1− control. In (c), the prey contains an empty expression vector (pEmpty) or a vector for the arabinose-inducible expression of Tmi1 (pTmi1). In (d), attackers contain an empty expression vector (pEmpty) or a vector for the arabinose-inducible expression of Tme1 (pTme1). Asterisks denote statistical significance between samples at the 4 h timepoint by an unpaired, two-tailed Student’s t-test (*P < 0.005; **P < 0.0005; ***P < 0.00005). Source data are provided as a source data file.
