Genome-Wide Identification by Transposon Insertion Sequencing of Escherichia coli K1 Genes Essential for in vitro Growth, Gastrointestinal Colonizing Capacity and Survival in Serum.


McCarthy, AJ; Stabler, RA; Taylor, PW; (2018) Genome-Wide Identification by Transposon Insertion Sequencing of Escherichia coli K1 Genes Essential for in vitro Growth, Gastrointestinal Colonizing Capacity and Survival in Serum. Journal of bacteriology. ISSN 0021-9193 DOI: https://doi.org/10.1128/JB.00698-17

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Abstract

Escherichia coli K1 strains are major causative agents of invasive disease of the new born. The age dependency of infection can be reproduced in the neonatal rat. Colonization of the small intestine following oral administration of K1 bacteria leads rapidly to invasion of the blood circulation; bacteria that avoid capture by the mesenteric lymphatic system and evade antibacterial mechanisms in the blood may disseminate to cause organ-specific infections such as meningitis. Some E. coli K1 surface constituents, in particular the polysialic acid capsule, are known to contribute to invasive potential but a comprehensive picture of the factors that determine the fully virulent phenotype has not so far emerged. We constructed a library and constituent sub-libraries of ∼775,000 Tn5 transposon mutants of E. coli K1 strain A192PP and employed transposon-directed insertion site sequencing (TraDIS) to identify genes required for fitness for infection in the two-day-old rat. Transposon insertions were lacking in 357 genes following recovery on selective agar; these genes were considered essential for growth in nutrient replete medium. Colonization of the mid-section of the small intestine was facilitated by 167 E. coli K1 gene products. Restricted bacterial translocation across epithelial barriers precluded TraDIS analysis of gut-to-blood and blood-to-brain transits; 97 genes were required for survival in human serum. The study revealed that a large number of bacterial genes, many not previously associated with systemic E. coli K1 infection, are required to realise full invasive potential.IMPORTANCEEscherichia coli K1 strains cause life-threatening infections in newborn infants. They are acquired from the mother at birth and colonize the small intestine, from where they invade the blood and central nervous system. It is difficult to obtain information from acutely ill patients that shed light on physiological and bacterial factors determining invasive disease. Key aspects of naturally occurring age-dependent human infection can be reproduced in neonatal rats. Here, we employ transposon-directed insertion site sequencing to identify genes essential for in vitro growth of E. coli K1 and genes that contribute to colonization of susceptible rats. The presence of bottlenecks to invasion of the blood and cerebrospinal compartments precluded insertion sequencing analysis but we identified genes for survival in serum.

Item Type: Article
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Pathogen Molecular Biology
Research Centre: Antimicrobial Resistance Centre (AMR)
PubMed ID: 29339415
Web of Science ID: 427113600011
URI: http://researchonline.lshtm.ac.uk/id/eprint/4646167

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