Pathway-Selective Sensitization of Mycobacterium tuberculosis for Target-Based Whole-Cell Screening

Garth L. Abrahams, Anuradha Kumar, Suzana Savvi, Alvin W. Hung, Shijun Wen, Chris Abell, Clifton E. Barry, David R. Sherman, Helena I.M. Boshoff, Valerie Mizrahi
2012 Chemistry and Biology  
Whole-cell screening of Mycobacterium tuberculosis (Mtb) remains a mainstay of drug discovery, but subsequent target elucidation often proves difficult. Conditional mutants that underexpress essential genes have been used to identify compounds with known mechanism of action by target-based wholecell screening (TB-WCS). Here, the feasibility of TB-WCS in Mtb was assessed by generating mutants that conditionally express pantothenate synthetase (panC), diaminopimelate decarboxylase (lysA), and
more » ... itrate lyase (icl1). The essentiality of panC and lysA, and conditional essentiality of icl1 for growth on fatty acids, was confirmed. Depletion of PanC and Icl1 rendered mutants hypersensitive to targetspecific inhibitors. Stable reporter strains were generated for use in high-throughput screening, and their utility was demonstrated by identifying compounds that display greater potency against a PanC-depleted strain. These findings illustrate the power of TB-WCS as a tool for tuberculosis drug discovery.
doi:10.1016/j.chembiol.2012.05.020 pmid:22840772 pmcid:PMC3421836 fatcat:sy5nt6drtndktfefumfpzej2ki