title: 14-3-3 proteins restrain the Exo1 nuclease to prevent overresection.
abstract: The DNA end resection process dictates the cellular response to DNA double strand break damage and is essential for genome maintenance. Although insufficient DNA resection hinders homology-directed repair and ATR (ataxia telangiectasia and Rad3 related)-dependent checkpoint activation, overresection produces excessive single-stranded DNA that could lead to genomic instability. However, the mechanisms controlling DNA end resection are poorly understood. Here we show that the major resection nuclease Exo1 is regulated both positively and negatively by protein-protein interactions to ensure a proper level of DNA resection. We have shown previously that the sliding DNA clamp proliferating cell nuclear antigen (PCNA) associates with the C-terminal domain of Exo1 and promotes Exo1 damage association and DNA resection. In this report, we show that 14-3-3 proteins interact with a central region of Exo1 and negatively regulate Exo1 damage recruitment and subsequent resection. 14-3-3s limit Exo1 damage association, at least in part, by suppressing its association with PCNA. Disruption of the Exo1 interaction with 14-3-3 proteins results in elevated sensitivity of cells to DNA damage. Unlike Exo1, the Dna2 resection pathway is apparently not regulated by PCNA and 14-3-3s. Our results provide critical insights into the mechanism and regulation of the DNA end resection process and may have implications for cancer treatment.
authors.affiliation: From the Departments of Cell Biology and Physiology and.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Chen
authors.affiliation: Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110 and.
authors.forename: In-Kwon
authors.initials: IK
authors.lastname: Kim
authors.affiliation: From the Departments of Cell Biology and Physiology and.
authors.forename: Yuchi
authors.initials: Y
authors.lastname: Honaker
authors.affiliation: From the Departments of Cell Biology and Physiology and.
authors.forename: Sharad C
authors.initials: SC
authors.lastname: Paudyal
authors.affiliation: From the Departments of Cell Biology and Physiology and.
authors.forename: Won Kyun
authors.initials: WK
authors.lastname: Koh
authors.affiliation: From the Departments of Cell Biology and Physiology and.
authors.forename: Melanie
authors.initials: M
authors.lastname: Sparks
authors.affiliation: From the Departments of Cell Biology and Physiology and.
authors.forename: Shan
authors.initials: S
authors.lastname: Li
authors.affiliation: From the Departments of Cell Biology and Physiology and the Department of Cancer Biology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77230.
authors.forename: Helen
authors.initials: H
authors.lastname: Piwnica-Worms
authors.affiliation: Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110 and.
authors.forename: Tom
authors.initials: T
authors.lastname: Ellenberger
authors.affiliation: From the Departments of Cell Biology and Physiology and zyou@wustl.edu.
authors.forename: Zhongsheng
authors.initials: Z
authors.lastname: You
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D004247:DNA / Q000235:genetics; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D014981:Xenopus
chemical_list: D048948:14-3-3 Proteins; D018809:Proliferating Cell Nuclear Antigen; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; D005982:Glutathione Transferase; D005090:Exodeoxyribonucleases; C028450:exodeoxyribonuclease I
keywords: 14-3-3 protein; DNA damage response; DNA repair; DNA resection; Exo1; checkpoint control; proliferating cell nuclear antigen (PCNA)
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