Abstract
Campylobacter jejuni is a major human pathogen and a leading cause of bacterial derived gastroenteritis worldwide. C. jejuni regulates gene expression under various environmental conditions and stresses, indicative of its ability to survive in diverse niches. Despite this ability to highly regulate gene transcription, C. jejuni encodes few transcription factors and its genome lacks many canonical transcriptional regulators. High throughput deep sequencing of mRNA transcripts (termed RNAseq) has been used to study the transcriptome of many different organisms, including C. jejuni; however, this technology has yet to be applied to defining the transcriptome of C. jejuni during in vivo colonization of its natural host, the chicken. In addition to its use in profiling the abundance of annotated genes, RNAseq is a powerful tool for identifying and quantifying, as-of-yet, unknown transcripts including non-coding regulatory RNAs, 5’ untranslated regulatory elements, and anti-sense transcripts. Here we report the complete transcriptome of C. jejuni during colonization of the chicken cecum and in two different in vitro growth phases using strand-specific RNAseq. Through this study, we identified over 250 genes differentially expressed in vivo in addition to numerous putative regulatory RNAs, including trans-acting non-coding RNAs and anti-sense transcripts. These latter potential regulatory elements were not identified in two prior studies using ORF-based microarrays, highlighting the power and value of the RNAseq approach. Our results provide new insights into how C. jejuni responds and adapts to the cecal environment and reveals new functions involved in colonization of its natural host.
Highlights
Campylobacter jejuni is an important human pathogen and a leading cause of bacterial derived gastroenteritis worldwide [1,2]
Much research has been carried out characterizing regulatory mechanisms in this human pathogen, most of this work has focused on in vitro studies with little knowledge generated towards understanding the mechanisms of global gene expression during colonization
Bar-coded cDNA libraries were generated from three independent cultures of C. jejuni grown in Mueller Hinton Broth (MHB) to mid-log and stationary phase, respectively
Summary
Campylobacter jejuni is an important human pathogen and a leading cause of bacterial derived gastroenteritis worldwide [1,2]. Much research has been carried out characterizing regulatory mechanisms in this human pathogen, most of this work has focused on in vitro studies with little knowledge generated towards understanding the mechanisms of global gene expression during colonization. C. jejuni highly regulates gene expression under various stresses and environmental conditions [4,12,13,14,15,16,17,18]. Despite its ability to rapidly alter gene expression, C. jejuni encodes only three sigma factors and a total of 34 other identified transcriptional regulators [9,10]. There appears to be a paucity of encoded transcription factors, other mechanisms of gene regulation have been identified in C
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