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

被引:43
作者
Ren, DC
Bedzyk, LA
Setlow, P
England, DF
Kjelleberg, S
Thomas, SM
Ye, RW
Wood, TK
机构
[1] Univ Connecticut, Dept Chem Engn, Storrs, CT 06269 USA
[2] Univ Connecticut, Dept Mol & Cell Biol, Storrs, CT 06269 USA
关键词
D O I
10.1128/AEM.70.8.4941-4949.2004
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
(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 mug of furanone per ml, while 5 mug of furanone per ml inhibited growth approximately two fold 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 jig 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 11 or TV 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 jig 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.
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页码:4941 / 4949
页数:9
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