DFI-seq identification of environment-specific gene expression in uropathogenic Escherichia coli

BMC Microbiology

Research Article

Tina Kronborg¹  Thomas Koed Doktor¹  Michelle Madelung¹  Jakob Møller-Jensen¹  Carsten Struve²  Karen Angeliki Krogfelt² 

[1] Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230, Odense M, Denmark;Department of Microbiology and Infection Control, Statens Serum Institut, Artillerivej 5, 2300, Copenhagen S, Denmark;

关键词: DFI;    NGS;    UPEC;    Amino acid biosynthesis;    Virulence;    UTI;   

DOI  :  10.1186/s12866-017-1008-4

 received in 2016-12-22, accepted in 2017-04-12,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundDuring infection of the urinary tract, uropathogenic Escherichia coli (UPEC) are exposed to different environments, such as human urine and the intracellular environments of bladder epithelial cells. Each environment elicits a distinct bacterial environment-specific transcriptional response. We combined differential fluorescence induction (DFI) with next-generation sequencing, collectively termed DFI-seq, to identify differentially expressed genes in UPEC strain UTI89 during growth in human urine and bladder cells.ResultsDFI-seq eliminates the need for iterative cell sorting of the bacterial library and yields a genome-wide view of gene expression. By analysing the gene expression of UPEC in human urine we found that genes involved in amino acid biosynthesis were upregulated. Deletion mutants lacking genes involved in arginine biosynthesis were outcompeted by the wild type during growth in human urine and inhibited in their ability to invade or proliferate in the J82 bladder epithelial cell line. Furthermore, DFI-seq was used to identify genes involved in invasion of J82 bladder epithelial cells. 56 genes were identified to be differentially expressed of which almost 60% encoded hypothetical proteins. One such gene UTI89_C5139, displayed increased adhesion and invasion of J82 cells when deleted from UPEC strain UTI89.ConclusionsWe demonstrate the usefulness of DFI-seq for identification of genes required for optimal growth of UPEC in human urine, as well as potential virulence genes upregulated during infection of bladder cell culture. DFI-seq holds potential for the study of bacterial gene expression in live-animal infection systems. By linking fitness genes, such as those genes involved in amino acid biosynthesis, to virulence, this study contributes to our understanding of UPEC pathophysiology.

【 授权许可】

CC BY   
© The Author(s). 2017