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Shigella species Gram-negative bacteria which cause a diarrheal disease, known as shigellosis, by invading and destroying the colonic mucosa and inducing a robust inflammatory response. With no vaccine available, shigellosis annually kills over 600,000 children in developing countries. This study demonstrates the utility of combining high-throughput bioinformatic methods within vitro andin vivo assays to provide new insights into pathogenesis. Comparisons ofin vivo andin vitro gene expression identified genes associated with intracellular growth. Additional bioinformatics analyses identified genes that are present inS. flexneri isolates but not in the three otherShigella species. Comparison of these two analyses revealed nine genes that are differentially expressed during invasion and that are specific toS. flexneri . One gene, a DeoR family transcriptional regulator with decreased expression during invasion, was further characterized and is now designatedicgR , fori ntrac ellularg rowthr egulator. Deletion oficgR caused no difference in growthin vitro but resulted in increased intracellular replication in HCT-8 cells. Furtherin vitro andin vivo studies using high-throughput sequencing of RNA transcripts (RNA-seq) of an isogenic ΔicgR mutant identified 34 genes that were upregulated under both growth conditions. This combined informatics and functional approach has allowed the characterization of a gene and pathway previously unknown inShigella pathogenesis and provides a framework for further identification of novel virulence factors and regulatory pathways.

Characterization of Intracellular Growth Regulator icgR by Utilizing Transcriptomics To Identify Mediators of Pathogenesis in Shigella flexneri