title: p53-Dependent DNA damage response sensitive to editing-defective tRNA synthetase in zebrafish.
abstract: Brain and heart pathologies are caused by editing defects of transfer RNA (tRNA) synthetases, which preserve genetic code fidelity by removing incorrect amino acids misattached to tRNAs. To extend understanding of the broader impact of synthetase editing reactions on organismal homeostasis, and based on effects in bacteria ostensibly from small amounts of mistranslation of components of the replication apparatus, we investigated the sensitivity to editing of the vertebrate genome. We show here that in zebrafish embryos, transient overexpression of editing-defective valyl-tRNA synthetase (ValRS(ED)) activated DNA break-responsive H2AX and p53-responsive downstream proteins, such as cyclin-dependent kinase (CDK) inhibitor p21, which promotes cell-cycle arrest at DNA damage checkpoints, and Gadd45 and p53R2, with pivotal roles in DNA repair. In contrast, the response of these proteins to expression of ValRS(ED) was abolished in p53-deficient fish. The p53-activated downstream signaling events correlated with suppression of abnormal morphological changes caused by the editing defect and, in adults, reversed a shortened life span (followed for 2 y). Conversely, with normal editing activities, p53-deficient fish have a normal life span and few morphological changes. Whole-fish deep sequencing showed genomic mutations associated with the editing defect. We suggest that the sensitivity of p53 to expression of an editing-defective tRNA synthetase has a critical role in promoting genome integrity and organismal homeostasis.
authors.affiliation: The Scripps Laboratories for tRNA Synthetase Research, The Scripps Research Institute, La Jolla, CA 92037; Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Youngzee
authors.initials: Y
authors.lastname: Song
authors.affiliation: The Scripps Laboratories for tRNA Synthetase Research, The Scripps Research Institute, La Jolla, CA 92037; Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
authors.forename: Yi
authors.initials: Y
authors.lastname: Shi
authors.affiliation: The Scripps Translational Science Institute, The Scripps Research Institute, La Jolla, CA 92037; Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037; Department of Human Biology, The J. Craig Venter Institute, La Jolla, CA 92037;
authors.forename: Tristan M
authors.initials: TM
authors.lastname: Carland
authors.affiliation: Department of Metabolism and Aging, The Scripps Research Institute, Jupiter, FL 33458;
authors.forename: Shanshan
authors.initials: S
authors.lastname: Lian
authors.affiliation: Department of Metabolism and Aging, The Scripps Research Institute, Jupiter, FL 33458;
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Sasaki
authors.affiliation: The Scripps Translational Science Institute, The Scripps Research Institute, La Jolla, CA 92037; Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037; Department of Human Biology, The J. Craig Venter Institute, La Jolla, CA 92037;
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Schork
authors.affiliation: Next Generation Sequencing and Microarray Core Facility, The Scripps Research Institute, La Jolla, CA 92037.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Head
authors.affiliation: Department of Metabolism and Aging, The Scripps Research Institute, Jupiter, FL 33458; kishi@scripps.edu schimmel@scripps.edu.
authors.forename: Shuji
authors.initials: S
authors.lastname: Kishi
authors.affiliation: The Scripps Laboratories for tRNA Synthetase Research, The Scripps Research Institute, La Jolla, CA 92037; Department of Cell and Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Metabolism and Aging, The Scripps Research Institute, Jupiter, FL 33458; kishi@scripps.edu schimmel@scripps.edu.
authors.forename: Paul
authors.initials: P
authors.lastname: Schimmel
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D004249:DNA Damage; D004625:Embryo, Nonmammalian / Q000196:embryology / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008297:Male; D009154:Mutation; D017393:RNA Editing; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D016159:Tumor Suppressor Protein p53; D029961:Zebrafish Proteins; D000604:Amino Acyl-tRNA Synthetases
keywords: genomic fidelity; genomic mutations; mistranslation; morphological changes; shortened lifespan