Whole-Genome Sequencing and iPLEX MassARRAY Genotyping Map an EMS-Induced Mutation Affecting Cell Competition in Drosophila melanogaster

G3 (Bethesda). 2016 Oct 13;6(10):3207-3217. doi: 10.1534/g3.116.029421.

Abstract

Cell competition, the conditional loss of viable genotypes only when surrounded by other cells, is a phenomenon observed in certain genetic mosaic conditions. We conducted a chemical mutagenesis and screen to recover new mutations that affect cell competition between wild-type and RpS3 heterozygous cells. Mutations were identified by whole-genome sequencing, making use of software tools that greatly facilitate the distinction between newly induced mutations and other sources of apparent sequence polymorphism, thereby reducing false-positive and false-negative identification rates. In addition, we utilized iPLEX MassARRAY for genotyping recombinant chromosomes. These approaches permitted the mapping of a new mutation affecting cell competition when only a single allele existed, with a phenotype assessed only in genetic mosaics, without the benefit of complementation with existing mutations, deletions, or duplications. These techniques expand the utility of chemical mutagenesis and whole-genome sequencing for mutant identification. We discuss mutations in the Atm and Xrp1 genes identified in this screen.

Keywords: Drosophila melanogaster; Flybook; Xrp1; cell competition; iPLEX MassARRAY; whole-genome sequencing.

Publication types

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

MeSH terms

  • Alleles
  • Animals
  • Ataxia Telangiectasia Mutated Proteins / genetics
  • Chromosome Mapping*
  • DNA Repair
  • Drosophila melanogaster / drug effects*
  • Drosophila melanogaster / genetics*
  • Ethyl Methanesulfonate / pharmacology*
  • Genetic Association Studies
  • Genetic Testing
  • Genome, Insect*
  • Genotype*
  • High-Throughput Nucleotide Sequencing*
  • Mutagenesis
  • Mutation / drug effects*
  • Phenotype
  • Quantitative Trait, Heritable
  • Recombination, Genetic

Substances

  • Ethyl Methanesulfonate
  • Ataxia Telangiectasia Mutated Proteins