A Staphylococcus aureus fitness test platform for mechanism-based profiling of antibacterial compounds.
The emergence of drug-resistant bacteria coupled with the limited discovery of novel chemical scaffolds and druggable targets inspires new approaches to antibiotic development. Here we describe a chemical genomics strategy based on 245 Staphylococcus aureus antisense RNA strains, each engineered for reduced expression of target genes essential for S. aureus growth. Attenuation of gene expression can sensitize cells to compounds that inhibit the activity of a gene product or associated process. Pools of strains grown competitively in the presence of bioactive compounds generate characteristic profiles of strain sensitivities reflecting compound mechanism of action. Here, we validate this approach with a structurally and mechanistically diverse set of reference antibiotics and, in the accompanying paper in this issue of Chemistry & Biology (Huber et al., 2009), demonstrate its use in the discovery of new cell wall inhibitors.
Donald
Robert G K
RG
Merck Research Laboratories, Merck & Co., Rahway, NJ 07065, USA.
Skwish
Stephen
S
Forsyth
R Allyn
RA
Anderson
Jennifer W
JW
Zhong
Tanya
T
Burns
Colleen
C
Lee
Suzy
S
Meng
Xin
X
LoCastro
Lynn
L
Jarantow
Lisa Wang
LW
Martin
Jesus
J
Lee
Sang Ho
SH
Taylor
Ian
I
Robbins
David
D
Malone
Cheryl
C
Wang
Liangsu
L
Zamudio
Carlos S
CS
Youngman
Philip J
PJ
Phillips
John W
JW
D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D016000:Cluster Analysis; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D010641:Phenotype; D034622:RNA Interference; D016372:RNA, Antisense / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D016372:RNA, Antisense