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BMC Microbiol ; 16(1): 130, 2016 06 27.
Artículo en Inglés | MEDLINE | ID: mdl-27349222

RESUMEN

BACKGROUND: Escherichia coli K-12 strains contain DNA cytosine methyltransferase (Dcm), which generates 5-methylcytosine at 5'CCWGG3' sites. Although the role of 5-methylcytosine in eukaryotic gene expression is relatively well described, the role of 5-methylcytosine in bacterial gene expression is largely unknown. RESULTS: To identify genes that are controlled by 5-methylcytosine in E. coli, we compared the transcriptomes of cells grown in the absence and presence of the DNA methylation inhibitor 5-azacytidine. We observed expression changes for 63 genes. The majority of the gene expression changes occurred at early stationary phase and were up-regulations. To identify gene expression changes due to a loss of DNA methylation, we compared the expression of selected genes in a wild-type and dcm knockout strain via reverse transcription quantitative PCR. CONCLUSIONS: Our data indicate that 5-azacytidine can influence gene expression by at least two distinct mechanisms: DNA methylation loss and a mechanism that is independent of DNA methylation loss. In addition, we have identified new targets of 5-methylcytosine-mediated regulation of gene expression. In summary, our data indicate that 5-azacytidine impacts the composition of the bacterial transcriptome, and the primary effect is increased gene expression at early stationary phase.


Asunto(s)
Azacitidina/antagonistas & inhibidores , Metilación de ADN/efectos de los fármacos , Escherichia coli K12/efectos de los fármacos , Escherichia coli K12/genética , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Transcriptoma/efectos de los fármacos , 5-Metilcitosina/metabolismo , 5-Metilcitosina/fisiología , Secuencia de Bases , Técnicas de Cultivo de Célula , Citosina , ADN Bacteriano , Escherichia coli K12/crecimiento & desarrollo , Escherichia coli K12/metabolismo , Proteínas de Escherichia coli/genética , Técnicas de Inactivación de Genes , Genes Bacterianos , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa , ARN Bacteriano/aislamiento & purificación , Análisis de Secuencia , Regulación hacia Arriba
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