Identification of inhibitors for single-stranded DNA-binding proteins in eubacteria.
The increasing threat of drug-resistant bacteria establishes a continuing need for the development of new strategies to fight infection. We examine the inhibition of the essential single-stranded DNA-binding proteins (SSBs) SSBA and SSBB as a potential antimicrobial therapy due to their importance in DNA replication, activating the SOS response and promoting competence-based mechanisms of resistance by incorporating new DNA.



Purified recombinant SSBs from Gram-positive (Staphylococcus aureus and Bacillus anthracis) and Gram-negative (Escherichia coli and Francisella tularensis) bacteria were assessed in a high-throughput screen for inhibition of duplex DNA unwinding by small molecule inhibitors. Secondary electrophoretic mobility shift assays further validated the top hits that were then tested for MICs using in vitro assays.



We have identified compounds that show cross-reactivity in vitro, as well as inhibition of both F. tularensis and B. anthracis SSBA. Five compounds were moderately toxic to at least two of the four bacterial strains in vivo, including two compounds that were selectively non-toxic to human cells, 9-hydroxyphenylfluoron and purpurogallin. Three of the SSBA inhibitors also inhibited S. aureus SSBB in Gram-positive bacteria.



Results from our study support the potential for SSB inhibitors as broad-spectrum antibacterial agents, with dual targeting capabilities against Gram-positive bacteria.
Department of Oral Biology, University of Nebraska Medical Center, Omaha, NE 68583, USA.|RPAcheQ, 16612 Martha Circle, Suite 510, Omaha, NE 68130, USA.
Jason G
JG
Glanzer
Department of Pathology and Microbiology, University of Nebraska Medical Center, 986495 Nebraska Medical Center, Omaha, NE 68198-6495, USA.
Jennifer L
JL
Endres
Department of Oral Biology, University of Nebraska Medical Center, Omaha, NE 68583, USA.
Brendan M
BM
Byrne
Department of Oral Biology, University of Nebraska Medical Center, Omaha, NE 68583, USA.
Shengqin
S
Liu
Department of Pathology and Microbiology, University of Nebraska Medical Center, 986495 Nebraska Medical Center, Omaha, NE 68198-6495, USA.
Kenneth W
KW
Bayles
Department of Oral Biology, University of Nebraska Medical Center, Omaha, NE 68583, USA goakley@unmc.edu.|Eppley Cancer Center, University of Nebraska Medical Center, Omaha, NE 68198, USA.
Greg G
GG
Oakley
D000900:Anti-Bacterial Agents / Q000302:isolation & purification* / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057166:High-Throughput Screening Assays; D008826:Microbial Sensitivity Tests
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins