A high proportion of these genes are regulated by SigA, which encodes the primary s factor of RNA buy 125-65-5 polymerase and is essential for cell growth. This result is in agreement with other studies on antibiotic treatment in B. subtilis, as there are some genes known to be induced by different antibiotics, such as yvgN, ywiE, pyrB, and purC. Although some characterized enzymes were present, including phosphoribosylglycinamide synthetase, many of the other genes encode proteins with no known function. The dlt operon, including dltA, is involved in the D-alanine esterification of lipoteichoic and wall teichoic acids, which increases bacterialMechanisms of Fusaricidins to Bacillus subtilisTable 2. Gene groups with E ,0.05 at 5, 20, and 170 min.Time point of fermentation: 5 min Gene groups SigW CcpA-negative SigK SigE AbrB-negative AbrB-positive GerE-negative FNR-positive SigBt value15.46 7.12 6.50 4.99 4.94 4.80 4.64 4.47 28.E value,10E-15 1.50E-09 1.12E-07 8.38E-04 1.09E-03 2.20E-03 4.83E-03 1.08E-02 9.25E-Mean 0.978 0.376 0.390 0.263 0.374 0.592 0.847 0.871 20.ORFs 63 120 93 148 61 21 10 9Time point of fermentation: 20 min Gene groups CcpA-negative SigK AbrB-positive StrCon-negative SigE SigH StrCon-positive PyrR-negative CtsR-negative PurR-negative SigBt value7.09 6.34 5.86 5.48 5.22 24.37 24.62 24.86 25.93 28.76 225.E value 1.86E-09 3.19E-07 6.43E-06 5.91E-05 2.49E-04 1.71E-02 5.32E-03 1.63E-03 4.21E-06 ,1.0E-15 ,1.0E-Mean 0.737 0.750 1.380 0.672 0.537 20.587 20.414 21.547 21.579 1.314 22.ORFs 120 93 21 92 148 32 59 9 12 32Time point of fermentation: 170 min Gene groups SigD Fur-negative SigA Ccp-negative CodY-negative StrCon-negative SinR-negative lolR-negative AbrB-positive SigL AbrB-negative Rok-negative SigBt value12.98 10.43 8.77 8.08 5.97 5.66 5.50 4.75 4.37 4.18 24.79 25.85 224.E value,1.0E-15 ,1.0E-15 ,1.0E-15 9.25E-13 3.29E-06 2.10E-05 5.28E-05 2.82E-03 1.71E-02 3.97E-02 2.31E-03 6.83E-06 ,1.0E-Mean 1.588 1.907 0.376 0.820 1.016 0.673 1.377 1.556 1.106 1.011 20.597 21.124 22.ORFs 77 36 674 120 44 92 21 13 21 23 61 29doi:10.1371/journal.pone.0050003.tresistance to cationic antimicrobial peptides. In this study, dltA was induced by more than 3-fold at 20 and 170 min (3.7- and 3.0-fold, respectively; Table S1), indicating that fusaricidin likely damages the cell wall and, in response, B. subtilis induces dltA. According to the MIPS analysis, the genes altered are mainly involved in glycolysis, the TCA cycle, and amino acid and fatty acid metabolisms (Fig. 4). CggR, YqzB, and YsiA were also significantlychanged by the fusaricidin treatment. CggR modulates 1407003 glucose catabolism, and it controls the expression of pgm, gapA, pgk, tpiA, and eno. pckA, sped, and gapB, all under the control of YqzB, were significantly repressed. Meanwhile, the genes regulated by the protein YsiA were overexpressed at 20 min, indicating that fusaricidin treatment may increase the degradation of fatty acids and histidine. As shown in Figure 4, the fusaricidin treatment MedChemExpress 298690-60-5 increased the catabolism of fatty acids and amino acids but strongly repressed glucose decomposition and gluconeogenesis. This phenomenon indicates that the strains require increased energy to mount defenses against antibiotic peptides. It is generally thought that if the culture medium lacks sugars, amino acids are then broken 1662274 down to provide the required carbon resource for other metabolic activities and gluconeogenesis should be concomitantly stimulated. However, our results appear to contrast with.A high proportion of these genes are regulated by SigA, which encodes the primary s factor of RNA polymerase and is essential for cell growth. This result is in agreement with other studies on antibiotic treatment in B. subtilis, as there are some genes known to be induced by different antibiotics, such as yvgN, ywiE, pyrB, and purC. Although some characterized enzymes were present, including phosphoribosylglycinamide synthetase, many of the other genes encode proteins with no known function. The dlt operon, including dltA, is involved in the D-alanine esterification of lipoteichoic and wall teichoic acids, which increases bacterialMechanisms of Fusaricidins to Bacillus subtilisTable 2. Gene groups with E ,0.05 at 5, 20, and 170 min.Time point of fermentation: 5 min Gene groups SigW CcpA-negative SigK SigE AbrB-negative AbrB-positive GerE-negative FNR-positive SigBt value15.46 7.12 6.50 4.99 4.94 4.80 4.64 4.47 28.E value,10E-15 1.50E-09 1.12E-07 8.38E-04 1.09E-03 2.20E-03 4.83E-03 1.08E-02 9.25E-Mean 0.978 0.376 0.390 0.263 0.374 0.592 0.847 0.871 20.ORFs 63 120 93 148 61 21 10 9Time point of fermentation: 20 min Gene groups CcpA-negative SigK AbrB-positive StrCon-negative SigE SigH StrCon-positive PyrR-negative CtsR-negative PurR-negative SigBt value7.09 6.34 5.86 5.48 5.22 24.37 24.62 24.86 25.93 28.76 225.E value 1.86E-09 3.19E-07 6.43E-06 5.91E-05 2.49E-04 1.71E-02 5.32E-03 1.63E-03 4.21E-06 ,1.0E-15 ,1.0E-Mean 0.737 0.750 1.380 0.672 0.537 20.587 20.414 21.547 21.579 1.314 22.ORFs 120 93 21 92 148 32 59 9 12 32Time point of fermentation: 170 min Gene groups SigD Fur-negative SigA Ccp-negative CodY-negative StrCon-negative SinR-negative lolR-negative AbrB-positive SigL AbrB-negative Rok-negative SigBt value12.98 10.43 8.77 8.08 5.97 5.66 5.50 4.75 4.37 4.18 24.79 25.85 224.E value,1.0E-15 ,1.0E-15 ,1.0E-15 9.25E-13 3.29E-06 2.10E-05 5.28E-05 2.82E-03 1.71E-02 3.97E-02 2.31E-03 6.83E-06 ,1.0E-Mean 1.588 1.907 0.376 0.820 1.016 0.673 1.377 1.556 1.106 1.011 20.597 21.124 22.ORFs 77 36 674 120 44 92 21 13 21 23 61 29doi:10.1371/journal.pone.0050003.tresistance to cationic antimicrobial peptides. In this study, dltA was induced by more than 3-fold at 20 and 170 min (3.7- and 3.0-fold, respectively; Table S1), indicating that fusaricidin likely damages the cell wall and, in response, B. subtilis induces dltA. According to the MIPS analysis, the genes altered are mainly involved in glycolysis, the TCA cycle, and amino acid and fatty acid metabolisms (Fig. 4). CggR, YqzB, and YsiA were also significantlychanged by the fusaricidin treatment. CggR modulates 1407003 glucose catabolism, and it controls the expression of pgm, gapA, pgk, tpiA, and eno. pckA, sped, and gapB, all under the control of YqzB, were significantly repressed. Meanwhile, the genes regulated by the protein YsiA were overexpressed at 20 min, indicating that fusaricidin treatment may increase the degradation of fatty acids and histidine. As shown in Figure 4, the fusaricidin treatment increased the catabolism of fatty acids and amino acids but strongly repressed glucose decomposition and gluconeogenesis. This phenomenon indicates that the strains require increased energy to mount defenses against antibiotic peptides. It is generally thought that if the culture medium lacks sugars, amino acids are then broken 1662274 down to provide the required carbon resource for other metabolic activities and gluconeogenesis should be concomitantly stimulated. However, our results appear to contrast with.
