Article Figures & Data
Figures
- Fig 1
Viable cell count of 18-h cultures. Stationary cultures of W3110 or PR200.1 were diluted to 5 × 105 CFU/ml in Mueller-Hinton broth and divided in three. (RFF)3RXB-FtsZ (FtsZ) or scrambled-base-sequence (Scr) control PPMO (160 μM) or no PPMO was added. Cultures were grown aerobically at 37°C for 18 h, and then samples of each were diluted and plated to determine viable cells. Error bars indicate standard deviations. *, highly significant (P < 0.01) difference compared to either the no-PPMO or scrambled-base-sequence (Scr) control-treated culture.
- Fig 2
Luciferase activity of cultures treated with PPMOs. Growing cultures were treated for 7 h without PPMO (no PPMO) or with 3 concentrations of a PPMO [(RFF)3RXB-Luc] targeted to a luciferase reporter gene or a scrambled-base-sequence control [(RFF)3RXB-Scr]. After 7 h, samples of each culture were measured for luciferase activity by luminometry. The experiment was repeated 3 times, and the error bars indicate standard deviations. *, a highly significant (P < 0.01) difference compared to cultures of PR200.1 with the same concentrations of (RFF)3RXB-Luc, the cultures of W3110 with the same concentrations of (RFF)3RXB-Scr, or the culture without PPMO.
Tables
- Table 1
MIC of AcpP PPMOs in pure cultures of E. coli
Motif, PPMO no.a Conjugated peptideb MIC (μM [μg/ml]) W3110 PR200.1 Motif 1 (C-N-N) NG-05-0200 RFFRFFRFFRXB 2.5 (14) 40 (222) NG-05-0653 DRDFDFDRDFDFDRDFDFDRXB 2.5 (14) 40 (222) NG-23-248 RXXRXXRXXRXB 20 (102) 80 (204) NG-06-0199 KFFKFFKFFKXB 10 (54) 80 (435) Motif 2 (C-N), NG-06-0073 RXRXRXRXRXRXB 1.25 (7) 1.25 (7) Motif 3 (C-N-C) NG-06-0076 RXRRXRRXRRXRXB 1.25 (7) 1.25 (7) NG-07-0795 RFRRFRRFRRFRXB 1 (6) 16 (94) No motif, NG-05-0246 RTRTRFLRRTXB 20 (111) 40 (111) - Table 2
MIC of (RFF)3R-AcpP (NG-05-0200) using oligopeptide transport mutants
E. coli strain Mutation/phenotype MIC μM μg/ml Morse 2034 Wild-type oligopeptide transport 5 28 PA0183 opp knockout, oligopeptide permease deletion 5 28 PA0333 dpp and opp knockout, PA0183 plus dipeptide permease deletion 5 28 PA0643 tpp, dpp, and opp knockout, PA0333 plus tripeptide permease mutant 5 28 - Table 3
MICs of standard antibiotics and PPMOs using transposome mutant RR3, isogenic parent strain XL1-Blue MRF′, and their sbmA complemented strains
PPMO no. Antibiotic or PPMO MIC (μM [μg/ml]) XL1-Blue MRF′ RR3 XL1-Blue MRF′ (pSbmA) RR3(pSbmA) Polymyxin B 0.8 (1) 0.8 (1) 0.8 (1) 0.8 (1) Colistin 0.9 (1) 0.9 (1) 0.9 (1) 0.9 (1) Erythromycin 34 (25) 34 (25) 34 (25) 34 (25) Rifampin 6 (5) 6 (5) 6 (5) 6 (5) Bleomycin 0.2 (0.25) 0.7 (1) 0.01 (0.016) 0.01 (0.016) Phleomycin 0.3 (0.5) 1.3 (2) 0.02 (0.03) 0.02 (0.03) NG-05-0200 (RFF)3RXB-AcpP 2 (11) 64 (355) 2 (11) 2 (11) NG-05-0653 [d-(RFF)3R]XB-AcpP 2 (11) 64 (355) 2 (11) 2 (11) NG-06-0076 (RXR)4XB-AcpP 1 (6) 8 (48) 0.1 (0.6) 0.1 (0.6) NG-05-0655 (RFF)3RXB-Scr >128 (>714) >128 (>714) >128 (>714) >128 (>714) NG-06-0078 (RXR)4XB-Scr >128 (>714) >128 (>714) >128 (>714) >128 (>714)
