title: Large-scale identification of chemically induced mutations in Drosophila melanogaster.
abstract: Forward genetic screens using chemical mutagens have been successful in defining the function of thousands of genes in eukaryotic model organisms. The main drawback of this strategy is the time-consuming identification of the molecular lesions causative of the phenotypes of interest. With whole-genome sequencing (WGS), it is now possible to sequence hundreds of strains, but determining which mutations are causative among thousands of polymorphisms remains challenging. We have sequenced 394 mutant strains, generated in a chemical mutagenesis screen, for essential genes on the Drosophila X chromosome and describe strategies to reduce the number of candidate mutations from an average of -3500 to 35 single-nucleotide variants per chromosome. By combining WGS with a rough mapping method based on large duplications, we were able to map 274 (-70%) mutations. We show that these mutations are causative, using small 80-kb duplications that rescue lethality. Hence, our findings demonstrate that combining rough mapping with WGS dramatically expands the toolkit necessary for assigning function to genes.
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Nele A
authors.initials: NA
authors.lastname: Haelterman
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA; Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Lichun
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA; Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Yumei
authors.initials: Y
authors.lastname: Li
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA; Medical Scientist Training Program, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Vafa
authors.initials: V
authors.lastname: Bayat
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Hector
authors.initials: H
authors.lastname: Sandoval
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Berrak
authors.initials: B
authors.lastname: Ugur
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Kai Li
authors.initials: KL
authors.lastname: Tan
authors.affiliation: Program in Structural and Computational Biology and Molecular Biophysics, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Ke
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Danqing
authors.initials: D
authors.lastname: Bei
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Bo
authors.initials: B
authors.lastname: Xiong
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Wu-Lin
authors.initials: WL
authors.lastname: Charng
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Theodore
authors.initials: T
authors.lastname: Busby
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Adeel
authors.initials: A
authors.lastname: Jawaid
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Gabriela
authors.initials: G
authors.lastname: David
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Manish
authors.initials: M
authors.lastname: Jaiswal
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA; Verna and Mars Mclean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
authors.forename: Koen J T
authors.initials: KJ
authors.lastname: Venken
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, Texas 77030, USA;
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA; Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas 77030, USA; hbellen@bcm.edu ruichen@bcm.edu.
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.affiliation: Program in Developmental Biology, Baylor College of Medicine, Houston, Texas 77030, USA; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA; Medical Scientist Training Program, Baylor College of Medicine, Houston, Texas 77030, USA; Program in Structural and Computational Biology and Molecular Biophysics, Baylor College of Medicine, Houston, Texas 77030, USA; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas 77030, USA; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, Texas 77030, USA; Department of Neuroscience, Baylor College of Medicine, Houston, Texas 77030, USA hbellen@bcm.edu ruichen@bcm.edu.
authors.forename: Hugo J
authors.initials: HJ
authors.lastname: Bellen
mesh_terms: D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D004331:Drosophila melanogaster / Q000235:genetics*; D005020:Ethyl Methanesulfonate; D005260:Female; D020043:Genes, Essential; D017344:Genes, Insect; D008297:Male; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009153:Mutagens; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods; D014960:X Chromosome
chemical_list: D009153:Mutagens; D005020:Ethyl Methanesulfonate