Abstract

BackgroundDespite comprehensive investigation, the Escherichia coli SOS response system is not yet fully understood. We have applied custom designed whole genome tiling arrays to measure UV invoked transcriptional changes in E. coli. This study provides a more complete insight into the transcriptome and the UV irradiation response of this microorganism.ResultsWe detected a number of novel differentially expressed transcripts in addition to the expected SOS response genes (such as sulA, recN, uvrA, lexA, umuC and umuD) in the UV treated cells. Several of the differentially expressed transcripts might play important roles in regulation of the cellular response to UV damage. We have predicted 23 novel small peptides from our set of detected non-gene transcripts. Further, three of the predicted peptides were cloned into protein expression vectors to test the biological activity. All three constructs expressed the predicted peptides, in which two of them were highly toxic to the cell. Additionally, a remarkably high overlap with previously in-silico predicted non-coding RNAs (ncRNAs) was detected. Generally we detected a far higher transcriptional activity than the annotation suggests, and these findings correspond with previous transcription mappings from E. coli and other organisms.ConclusionsHere we demonstrate that the E. coli transcriptome consists of far more transcripts than the present annotation suggests, of which many transcripts seem important to the bacterial stress response. Sequence alignment of promoter regions suggest novel regulatory consensus sequences for some of the upregulated genes. Finally, several of the novel transcripts identified in this study encode putative small peptides, which are biologically active.

Highlights

  • UV irradiation is one of the most common treatments used to study cellular responses to DNA damage

  • For two of the upregulated non-coding RNAs (ncRNAs), istR-2 and rprA, no expression above the microarray background signal can be detected in the reference transcriptome

  • It is tempting to speculate that these two ncRNA can be direct targets of the SOS response pathway but this remains to be shown

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Summary

Introduction

UV irradiation is one of the most common treatments used to study cellular responses to DNA damage. In Escherichia coli UV irradiation transiently blocks DNA replication and leads to the induction of a physiological response termed the SOS response [1]. More than 40 genes induced by the SOS response are negatively regulated by the LexA repressor which binds to operator sequences upstream of genes or operons [2]. After UV irradiation RecA protein binds to ssDNA formed by a replication block, leading to the generation of a RecA-ssDNA nucleoprotein filament that is able to mediate auto-proteolysis of the LexA repressor. E. coli microarrays, carrying 95.5% of all annotated open reading frames, were utilized to identify changes in gene expression after UV irradiation [3]. This study provides a more complete insight into the transcriptome and the UV irradiation response of this microorganism

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