Clerocidin selectively modifies the gyrase-DNA gate to induce irreversible and reversible DNA damage.

Nucleic Acids Research
Xiao Su PanL Mark Fisher

Abstract

Clerocidin (CL), a microbial diterpenoid, reacts with DNA via its epoxide group and stimulates DNA cleavage by type II DNA topoisomerases. The molecular basis of CL action is poorly understood. We establish by genetic means that CL targets DNA gyrase in the gram-positive bacterium Streptococcus pneumoniae, and promotes gyrase-dependent single- and double-stranded DNA cleavage in vitro. CL-stimulated DNA breakage exhibited a strong preference for guanine preceding the scission site (-1 position). Mutagenesis of -1 guanines to A, C or T abrogated CL cleavage at a strong pBR322 site. Surprisingly, for double-strand breaks, scission on one strand consistently involved a modified (piperidine-labile) guanine and was not reversed by heat, salt or EDTA, whereas complementary strand scission occurred at a piperidine-stable -1 nt and was reversed by EDTA. CL did not induce cleavage by a mutant gyrase (GyrA G79A) identified here in CL-resistant pneumococci. Indeed, mutations at G79 and at the neighbouring S81 residue in the GyrA breakage-reunion domain discriminated poisoning by CL from that of antibacterial quinolones. The results suggest a novel mechanism of enzyme inhibition in which the -1 nt at the gyrase-DNA gate exhibit different C...Continue Reading

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Citations

Sep 10, 2011·Applied Microbiology and Biotechnology·Frédéric CollinAnthony Maxwell
Aug 14, 2013·Nucleic Acids Research·Elisa ArnoldiL Mark Fisher
Aug 5, 2015·Antimicrobial Agents and Chemotherapy·George A JacobyDavid C Hooper
Feb 20, 2009·Organic & Biomolecular Chemistry·Sara N RichterManlio Palumbo
Oct 6, 2015·EcoSal Plus·Natassja G BushAnthony Maxwell

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