title: Identification of suppressors of mbk-2/DYRK by whole-genome sequencing.
abstract: Screening for suppressor mutations is a powerful method to isolate genes that function in a common pathway or process. Because suppressor mutations often do not have phenotypes on their own, cloning of suppressor loci can be challenging. A method combining whole-genome sequencing (WGS) and single nucleotide polymorphism (SNP) mapping (WGS/SNP mapping) was developed to identify mutations with visible phenotypes in C. elegans. We show here that WGS/SNP mapping is an efficient method to map suppressor mutations without the need for previous phenotypic characterization. Using RNA-mediated interference to test candidate loci identified by WGS/SNP mapping, we identified 10 extragenic and six intragenic suppressors of mbk-2, a DYRK family kinase required for the transition from oocyte to zygote. Remarkably, seven suppressors are mutations in cell-cycle regulators that extend the timing of the oocyte-to-zygote transition.
authors.affiliation: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors.forename: Yuemeng
authors.initials: Y
authors.lastname: Wang
authors.forename: Jennifer T
authors.initials: JT
authors.lastname: Wang
authors.forename: Dominique
authors.initials: D
authors.lastname: Rasoloson
authors.forename: Michael L
authors.initials: ML
authors.lastname: Stitzel
authors.forename: Kevin F
authors.initials: KF
authors.lastname: O' Connell
authors.forename: Harold E
authors.initials: HE
authors.lastname: Smith
authors.forename: Geraldine
authors.initials: G
authors.lastname: Seydoux
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D020134:Catalytic Domain; D004843:Epistasis, Genetic; D018507:Gene Expression Regulation, Developmental; D049751:Genome, Helminth; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D017422:Sequence Analysis, DNA
chemical_list: D029742:Caenorhabditis elegans Proteins; C478059:MBK-2 protein, C elegans; D011505:Protein-Tyrosine Kinases
keywords: C. elegans; DYRK kinase; MBK-2; single nucleotide polymorphism mapping; suppressors; whole-genome sequencing