title: HCoDES reveals chromosomal DNA end structures with single-nucleotide resolution.
abstract: The structure of broken DNA ends is a critical determinant of the pathway used for DNA double-strand break (DSB) repair. Here, we develop an approach involving the hairpin capture of DNA end structures (HCoDES), which elucidates chromosomal DNA end structures at single-nucleotide resolution. HCoDES defines structures of physiologic DSBs generated by the RAG endonuclease, as well as those generated by nucleases widely used for genome editing. Analysis of G1 phase cells deficient in H2AX or 53BP1 reveals DNA ends that are frequently resected to form long single-stranded overhangs that can be repaired by mutagenic pathways. In addition to 3' overhangs, many of these DNA ends unexpectedly form long 5' single-stranded overhangs. The divergence in DNA end structures resolved by HCoDES suggests that H2AX and 53BP1 may have distinct activities in end protection. Thus, the high-resolution end structures obtained by HCoDES identify features of DNA end processing during DSB repair.
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Yair
authors.initials: Y
authors.lastname: Dorsett
authors.affiliation: Department of Pediatrics, Washington University School of Medicine, St. Louis, MO 63110, USA; Genome Institute, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Yanjiao
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Tubbs
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Bo-Ruei
authors.initials: BR
authors.lastname: Chen
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Caitlin
authors.initials: C
authors.lastname: Purman
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Baeck-Seung
authors.initials: BS
authors.lastname: Lee
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Rosmy
authors.initials: R
authors.lastname: George
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Bredemeyer
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Jiang-Yang
authors.initials: JY
authors.lastname: Zhao
authors.affiliation: Genome Institute, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Erica
authors.initials: E
authors.lastname: Sodergen
authors.affiliation: Genome Institute, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: George M
authors.initials: GM
authors.lastname: Weinstock
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Nathan D
authors.initials: ND
authors.lastname: Han
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Reyes
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Eugene M
authors.initials: EM
authors.lastname: Oltz
authors.affiliation: Biochemistry Department, St. Louis University School of Medicine, St. Louis, MO 63104, USA.
authors.forename: Dale
authors.initials: D
authors.lastname: Dorsett
authors.affiliation: Biochemistry Department, St. Louis University School of Medicine, St. Louis, MO 63104, USA.
authors.forename: Ziva
authors.initials: Z
authors.lastname: Misulovin
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA. Electronic address: jpayton@wustl.edu.
authors.forename: Jacqueline E
authors.initials: JE
authors.lastname: Payton
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA. Electronic address: sleckman@immunology.wustl.edu.
authors.forename: Barry P
authors.initials: BP
authors.lastname: Sleckman
mesh_terms: D001483:Base Sequence; D002478:Cells, Cultured; D002877:Chromosomes, Human / Q000235:genetics*; D053903:DNA Breaks, Double-Stranded; D006801:Humans; D017422:Sequence Analysis, DNA / Q000379:methods*