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Cell‐rounding compounds inhibit components of the bacterial cytoskeleton
Author(s) -
Lammers Matthew,
Santos Thiago,
Hurley Katherine,
Mitchinson Timothy,
Weibel Douglas
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.612.8
Subject(s) - mreb , cytoskeleton , actin , microbiology and biotechnology , cell division , biology , mutant , phenotype , actin cytoskeleton , cell , morphogenesis , chemistry , genetics , gene
The bacterial cytoskeleton is a highly organized complex consisting of actin, tubulin, and intermediate filament protein homologs. Particularly, the actin homolog MreB is important for chromosome segregation, cell wall morphogenesis, and maintenance of rod‐shape morphology. A22 is a small molecule bacterial inhibitor of MreB that interferes with normal growth and produces rounded cells. In this study, we identify the mechanism of six novel bacterial inhibitors that produce a similar rounded phenotype with varying degrees of chemical similarity to A22. We characterize the activity of these cell‐rounding compounds (CRCs) against diverse bacterial species and perform toxicity testing in mammalian cells to test the viability of these compounds as therapeutic agents. Furthermore, we characterize phenotypic aberrations by quantifying cellular aspect ratio and localization of MreB in treated cells via flow cytometry and structured illumination microscopy, respectively. To test the hypothesis that the CRCs bind the MreB ATPase catalytic center in a similar fashion to A22, we test A22‐resistant mutants for cross‐resistance. We also perform a multi‐copy suppression screen to determine cytoskeleton gene products which confer reduced susceptibility to these compounds and generate spontaneous mutants to identify mutations which provide resistance. Finally, we report a convenient chemical synthesis for the CRCs. This study will help identify the mechanism of action of new bioactive molecules and serve as a starting point for the development of a novel class of therapeutic agents.