TABLE 3.
Effect of genes whose inactivation prevented repression of hdeAB by MarA on MarA-mediated MDR
Protein/function | Gene | Locusb | Level of MarA-mediated MDR (% of parental level)a |
|||
---|---|---|---|---|---|---|
FOX | NOR | CHL | MIN | |||
Transcriptional regulators | ||||||
Cold shock protein CspG | cspGpc | b0990 | 54 | 91 | 93 | 75 |
Cyclic AMP receptor protein | crp | b3357 | 21 | 33 | 29 | 38 |
DNA-binding protein H-NS | hns | b1237 | 19 | 23 | 20 | 35 |
OmpR transcriptional dual regulator | ompR | b3405 | 67 | 77 | 93 | 100 |
Enzymes | ||||||
Acid phosphatase | appA | b0980 | 68 | 84 | 93 | 100 |
Adenylate cyclase | cyaA | b3806 | 43 | 41 | 46 | 32 |
Serine protease Do | degP | b0161 | 94 | 91 | 50 | 100 |
Putative ring-cleavage enzyme of phenylacetate degradation | maoC | b1387 | 100 | 61 | 75 | 40 |
Aspartate kinase/homoserine dehydrogenase | metL | b3940 | 120 | 91 | 100 | 100 |
Poly(A) polymerase I; RNA modification | pcnB | b0143 | 10 | 23 | 33 | 30 |
RecBCD DNA helicase/exonuclease | recD | b2819 | 80 | 77 | 50 | 65 |
Trehalose-6-phosphate hydrolase | treC | b4239 | 94 | 46 | 85 | 100 |
Oxidoreductase subunit paralog of DmsA | ynfE | b1587 | 94 | 68 | 93 | 100 |
Transport | ||||||
AcrAB-TolC multidrug efflux system | acrA | b0463 | 16 | 0 | 8 | 40 |
AcrAB-TolC multidrug efflux system | acrB | b0462 | 6 | 9 | 8 | 40 |
Putative propionic acid transporter | mhpT | b0353 | 68 | 91 | 100 | 100 |
Subunit of nickel ABC transporter | nikD | b3479 | 68 | 68 | 50 | 125 |
TolC outer membrane channel | tolC | b3035 | 0 | 0 | 0 | 40 |
Others | ||||||
Putative membrane protein | alx | b3088 | 120 | 68 | 100 | 65 |
Predicted membrane-anchored protein | damX | b3388 | 40 | 46 | 50 | 30 |
tRNA-methionine | metV | b2816 | 74 | 91 | 75 | 65 |
CPS-53 (KpLE1) prophage; bactoprenol-linked glucose translocase (flippase) | yfdG | b2350 | 68 | 91 | 93 | 65 |
Conserved protein | yibLpc | b3602 | 68 | 68 | 50 | 100 |
Predicted unconserved protein | yniDpc | b4535 | 94 | 77 | 125 | 50 |
Abbreviations: FOX, cefoxitin; NOR, norfloxacin; CHL, chloramphenicol; MIN, minocycline. MarA-mediated MDR for each mutant was calculated as the fold increase in MIC for the ΔmarR background (CR1000-derivative mutant) compared to the MIC for the ΔmarRA background (CR2000-derivative mutant) by using the ΔmarR MIC/ΔmarRA MIC ratio. This ratio for the parental strains (the MIC for the ΔmarR parental strain CR1000 divided by the MIC for the ΔmarRA parental strain CR2000) for each antibiotic (shown in Table 2, footnote b) was considered to represent 100% MarA-mediated antibiotic resistance, whereas a ratio of 1 (with the MIC for the ΔmarR background being equal to the MIC for the ΔmarRA background) was considered to represent 0% MarA-mediated antibiotic resistance. These two reference points were used to calculate the percentage of MarA-mediated antibiotic resistance for each mutant according to its ΔmarR MIC/ΔmarRA MIC ratio. When inactivation of a gene significantly reduced MarA-mediated resistance to an antibiotic compared to the level for the parental strain (P < 0.01, associated with reductions of 50% or more), the corresponding value is highlighted in bold. For parental strains, n = 9; for mutants, n = 2 to 5.
The locus tags refer to accession numbers (http://biocyc.org/ecocyc/index.shtml).
The insertion of the transposome was in the promoter of the gene. All the other insertions were within the open reading frame (ORF) of the genes.