Genome-wide detection of fitness genes in uropathogenic Escherichia coli during systemic infection

PLoS Pathogens
Sargurunathan SubashchandraboseHarry L T Mobley

Abstract

Uropathogenic Escherichia coli (UPEC) is a leading etiological agent of bacteremia in humans. Virulence mechanisms of UPEC in the context of urinary tract infections have been subjected to extensive research. However, understanding of the fitness mechanisms used by UPEC during bacteremia and systemic infection is limited. A forward genetic screen was utilized to detect transposon insertion mutants with fitness defects during colonization of mouse spleens. An inoculum comprised of 360,000 transposon mutants in the UPEC strain CFT073, cultured from the blood of a patient with pyelonephritis, was used to inoculate mice intravenously. Transposon insertion sites in the inoculum (input) and bacteria colonizing the spleen (output) were identified using high-throughput sequencing of transposon-chromosome junctions. Using frequencies of representation of each insertion mutant in the input and output samples, 242 candidate fitness genes were identified. Co-infection experiments with each of 11 defined mutants and the wild-type strain demonstrated that 82% (9 of 11) of the tested candidate fitness genes were required for optimal fitness in a mouse model of systemic infection. Genes involved in biosynthesis of poly-N-acetyl glucosamine (pg...Continue Reading

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Citations

Dec 10, 2014·Proceedings of the National Academy of Sciences of the United States of America·Sargurunathan SubashchandraboseHarry L T Mobley
Nov 1, 2015·Journal of Medical Microbiology·Helen MiajlovicStephen Gj Smith
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Datasets Mentioned

BETA
SRP027190
SRA

Methods Mentioned

BETA
Illumina sequencing
PCR
RNAseq
motility assay

Software Mentioned

BOWTIE
TFAST
TraDIS
Prism
INSeq
RNAseq
BLAST

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