Detection of the cell wall-affecting antibiotics at sublethal concentrations using a reporter Staphylococcus aureus harboring drp35 promoter - lacZ transcriptional fusion.
Previously, various inhibitors of cell wall synthesis induced the drp35 gene of Staphylococcus aureus efficiently. To determine whether drp35 could be exploited in antistaphylococcal drug discovery, we cloned the promoter of drp35 (P(d)) and developed different biological assay systems using an engineered S. aureus strain that harbors a chromosomally-integrated P(d) - lacZ transcriptional fusion. An agarose-based assay showed that P(d) is induced not only by the cell wall-affecting antibiotics but also by rifampicin and ciprofloxacin. In contrast, a liquid medium-based assay revealed the induction of P(d) specifically by the cell wall-affecting antibiotics. Induction of P(d) by sublethal concentrations of cell wall-affecting antibiotics was even assessable in a microtiter plate assay format, indicating that this assay system could be potentially used for high-throughput screening of new cell wall-inhibiting compounds.
Mondal
Rajkrishna
R
Department of Biochemistry, Bose Institute, Kolkata, West Bengal, India.
Chanda
Palas K
PK
Bandhu
Amitava
A
Jana
Biswanath
B
Lee
Chia Y
CY
Sau
Subrata
S
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002473:Cell Wall / Q000187:drug effects*; D002939:Ciprofloxacin / Q000494:pharmacology; D057166:High-Throughput Screening Assays; D006867:Hydrolases; D008826:Microbial Sensitivity Tests; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; C400364:drp35 protein, Staphylococcus aureus; D002939:Ciprofloxacin; D006867:Hydrolases; D001616:beta-Galactosidase; D012293:Rifampin