Haploid genetic screens in human cells identify host factors used by pathogens

Science. 2009 Nov 27;326(5957):1231-5. doi: 10.1126/science.1178955.

Abstract

Loss-of-function genetic screens in model organisms have elucidated numerous biological processes, but the diploid genome of mammalian cells has precluded large-scale gene disruption. We used insertional mutagenesis to develop a screening method to generate null alleles in a human cell line haploid for all chromosomes except chromosome 8. Using this approach, we identified host factors essential for infection with influenza and genes encoding important elements of the biosynthetic pathway of diphthamide, which are required for the cytotoxic effects of diphtheria toxin and exotoxin A. We also identified genes needed for the action of cytolethal distending toxin, including a cell-surface protein that interacts with the toxin. This approach has both conceptual and practical parallels with genetic approaches in haploid yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / metabolism
  • ADP Ribose Transferases / toxicity
  • Adenosine Diphosphate Ribose / metabolism
  • Amino Acid Sequence
  • Antigens, Bacterial / metabolism
  • Antigens, Bacterial / toxicity
  • Bacterial Toxins / metabolism*
  • Bacterial Toxins / toxicity
  • Biosynthetic Pathways
  • Carboxylic Ester Hydrolases
  • Cell Line, Tumor
  • Diphtheria Toxin / metabolism
  • Diphtheria Toxin / toxicity
  • Exotoxins / metabolism
  • Exotoxins / toxicity
  • Genes
  • Genetic Testing*
  • Haploidy*
  • Histidine / analogs & derivatives
  • Histidine / biosynthesis
  • Host-Pathogen Interactions*
  • Humans
  • Influenza A Virus, H1N1 Subtype / pathogenicity*
  • Molecular Sequence Data
  • Monosaccharide Transport Proteins / genetics
  • Monosaccharide Transport Proteins / metabolism
  • Mutagenesis, Insertional
  • N-Acylneuraminate Cytidylyltransferase / genetics
  • N-Acylneuraminate Cytidylyltransferase / metabolism
  • Peptide Elongation Factor 2 / metabolism
  • Proteins / chemistry
  • Proteins / genetics
  • Proteins / metabolism
  • Virulence Factors / metabolism
  • Virulence Factors / toxicity

Substances

  • Antigens, Bacterial
  • Bacterial Toxins
  • Diphtheria Toxin
  • Exotoxins
  • Monosaccharide Transport Proteins
  • Peptide Elongation Factor 2
  • Proteins
  • UDP-galactose translocator
  • Virulence Factors
  • anthrax toxin
  • cytolethal distending toxin
  • Adenosine Diphosphate Ribose
  • Histidine
  • diphthamide
  • DPH7 protein, human
  • ADP Ribose Transferases
  • toxA protein, Pseudomonas aeruginosa
  • N-Acylneuraminate Cytidylyltransferase
  • Carboxylic Ester Hydrolases