Gene Fitness Landscapes of Vibrio Cholerae at Important Stages of Its Life Cycle

PLoS Pathog. 2013;9(12):e1003800. doi: 10.1371/journal.ppat.1003800. Epub 2013 Dec 26.

Abstract

Vibrio cholerae has evolved to adeptly transition between the human small intestine and aquatic environments, leading to water-borne spread and transmission of the lethal diarrheal disease cholera. Using a host model that mimics the pathology of human cholera, we applied high density transposon mutagenesis combined with massively parallel sequencing (Tn-seq) to determine the fitness contribution of >90% of all non-essential genes of V. cholerae both during host infection and dissemination. Targeted mutagenesis and validation of 35 genes confirmed our results for the selective conditions with a total false positive rate of 4%. We identified 165 genes never before implicated for roles in dissemination that reside within pathways controlling many metabolic, catabolic and protective processes, from which a central role for glycogen metabolism was revealed. We additionally identified 76 new pathogenicity factors and 414 putatively essential genes for V. cholerae growth. Our results provide a comprehensive framework for understanding the biology of V. cholerae as it colonizes the small intestine, elicits profuse secretory diarrhea, and disseminates into the aquatic environment.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptation, Physiological / genetics
  • Animals
  • Animals, Newborn
  • Genetic Fitness*
  • Genome, Bacterial
  • High-Throughput Nucleotide Sequencing
  • Host-Pathogen Interactions
  • Life Cycle Stages / genetics*
  • Microbial Viability / genetics
  • Organisms, Genetically Modified
  • Rabbits
  • Vibrio cholerae / genetics*
  • Vibrio cholerae / growth & development*
  • Water

Substances

  • Water