Characterizing septum inhibition in Mycobacterium tuberculosis for novel drug discovery

A temperature sensitive mutation in the cell, division protein FtsZ was used in combination with transcriptional analysis to identify biomarkers for inhibition of septum formation. Crystallography and modeling revealed that the glycine for aspartate substitution at amino acid 210 was located in helix 8 of the protein, adjacent to the T7 synergy loop. To verify the molecular behavior of FtSZ(D210G), the in vitro activity and structural stability were evaluated as a function of temperature. These analyses confirmed that the FtsZ(D21OG) mutant had reduced GTPase and polymerization activity compared to wild-type FtsZ, and CD spectroscopy demonstrated that both FtsZD21OG and wild-type FtsZ had similar structure and stability. Significantly, the FtsZD21OG merodiploid strain of A tuberculosis had compromised growth at 37 degrees C, substantiating the suitability of FtSZ(D21OG) as a molecular toot for global analysis in response to improper FtsZ polymerization and septum inhibition. Advanced model-based bioinformatics and transcriptional mapping were used to identify high-content multiple features that provide biomarkers for the development of a rational drug screening platform for discovering novel chemotherapeutics that target cell division. (c) 2008 Published by Elsevier Ltd.