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Differential gene expression to investigate the effect of (5Z)-4-bromo- 5-(bromomethylene)-3-butyl-2(5H)-furanone on Bacillus subtilis.

Abstract
(5Z)-4-Bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone (furanone) from the red marine alga Delisea pulchra was found previously to inhibit the growth, swarming, and biofilm formation of gram-positive bacteria. Using the gram-positive bacterium Bacillus subtilis as a test organism, we observed cell killing by 20 microg of furanone per ml, while 5 microg of furanone per ml inhibited growth approximately twofold without killing the cells. To discover the mechanism of this inhibition on a genetic level and to investigate furanone as a novel antibiotic, full-genome DNA microarrays were used to analyze the gene expression profiles of B. subtilis grown with and without 5 microg of furanone per ml. This agent induced 92 genes more than fivefold (P < 0.05) and repressed 15 genes more than fivefold (P < 0.05). The induced genes include genes involved in stress responses (such as the class III heat shock genes clpC, clpE, and ctsR and the class I heat shock genes groES, but no class II or IV heat shock genes), fatty acid biosynthesis, lichenan degradation, transport, and metabolism, as well as 59 genes with unknown functions. The microarray results for four genes were confirmed by RNA dot blotting. Mutation of a stress response gene, clpC, caused B. subtilis to be much more sensitive to 5 microg of furanone per ml (there was no growth in 8 h, while the wild-type strain grew to the stationary phase in 8 h) and confirmed the importance of the induction of this gene as identified by the microarray analysis.
AuthorsDacheng Ren, Laura A Bedzyk, Peter Setlow, Dacre F England, Staffan Kjelleberg, Stuart M Thomas, Rick W Ye, Thomas K Wood
JournalApplied and environmental microbiology (Appl Environ Microbiol) Vol. 70 Issue 8 Pg. 4941-9 (Aug 2004) ISSN: 0099-2240 [Print] United States
PMID15294834 (Publication Type: Journal Article)
Chemical References
  • 4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone
  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Culture Media
  • Furans
Topics
  • Anti-Bacterial Agents (pharmacology)
  • Bacillus subtilis (drug effects, growth & development)
  • Bacterial Proteins (genetics, metabolism)
  • Culture Media
  • Furans (pharmacology)
  • Gene Expression Profiling
  • Gene Expression Regulation, Bacterial
  • Microscopy, Confocal
  • Oligonucleotide Array Sequence Analysis

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