Department of Biology, State University of New York at Geneseo, Geneseo, NY, USA.
FEMS Microbiol Lett. 2014 Jan;350(1):100-6. doi: 10.1111/1574-6968.12299. Epub 2013 Nov 6.
Escherichia coli K-12 strains contain the orphan cytosine-5 DNA methyltransferase enzyme Dcm (DNA cytosine methyltransferase). Two recent reports indicate that Dcm has an influence on stationary phase gene expression in E. coli. Herein, we demonstrate that dcm knockout cells overexpress the drug resistance transporter SugE, which has been linked to ethidium bromide (ETBR) resistance. SugE expression also increased in the presence of the DNA methylation inhibitor 5-azacytidine, suggesting that Dcm-mediated DNA methylation normally represses sugE expression. The effect of Dcm on sugE expression is primarily restricted to early stationary phase, and RpoS is required for robust sugE expression. Dcm knockout cells are more resistant to ETBR than wild-type cells, and complementation with a plasmid-borne dcm gene restores ETBR sensitivity. SugE knockout cells are more sensitive to ETBR than wild-type cells. These data indicate that Dcm influences the sensitivity to an antimicrobial compound through changes in gene expression.
大肠杆菌 K-12 菌株含有孤儿胞嘧啶-5 DNA 甲基转移酶酶 Dcm(DNA 胞嘧啶甲基转移酶)。最近有两项报告表明,Dcm 对大肠杆菌的静止期基因表达有影响。本文中,我们证明了 dcm 敲除细胞过表达了耐药转运蛋白 SugE,该蛋白与溴化乙锭(ETBR)耐药性有关。在 DNA 甲基化抑制剂 5-氮杂胞苷存在的情况下,SugE 的表达也增加了,这表明 Dcm 介导的 DNA 甲基化通常会抑制 sugE 的表达。Dcm 对 sugE 表达的影响主要局限于早期静止期,并且 RpoS 是强烈表达 sugE 所必需的。与野生型细胞相比,dcm 敲除细胞对 ETBR 的抗性更强,并且通过质粒携带的 dcm 基因的互补恢复了 ETBR 敏感性。与野生型细胞相比,SugE 敲除细胞对 ETBR 更敏感。这些数据表明,Dcm 通过改变基因表达来影响对抗微生物化合物的敏感性。