1. Phosphoproteomic analysis reveals plant DNA damage signalling pathways with a functional role for histone H2AX phosphorylation in plant growth under genotoxic stress.
- Author
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Waterworth WM, Wilson M, Wang D, Nuhse T, Warward S, Selley J, and West CE
- Subjects
- ATP-Binding Cassette Transporters genetics, Aging metabolism, Aging physiology, Aging radiation effects, Arabidopsis growth & development, Arabidopsis radiation effects, Arabidopsis Proteins genetics, Cells, Cultured, DNA Damage genetics, DNA Damage radiation effects, Gene Expression Regulation, Plant genetics, Gene Expression Regulation, Plant radiation effects, Gene Ontology, Germination genetics, Germination radiation effects, Histones chemistry, Mass Spectrometry, Phosphorylation, Proteome genetics, Proteome radiation effects, Seeds metabolism, Seeds physiology, Seeds radiation effects, Serine chemistry, Serine metabolism, Signal Transduction radiation effects, X-Rays, ATP-Binding Cassette Transporters metabolism, Arabidopsis metabolism, Arabidopsis Proteins metabolism, DNA Repair genetics, DNA Repair radiation effects, Histones metabolism, Proteome metabolism, Seeds growth & development, Stress, Physiological genetics, Stress, Physiological radiation effects
- Abstract
DNA damage responses are crucial for plant growth under genotoxic stress. Accumulating evidence indicates that DNA damage responses differ between plant cell types. Here, quantitative shotgun phosphoproteomics provided high-throughput analysis of the DNA damage response network in callus cells. MS analysis revealed a wide network of highly dynamic changes in the phosphoprotein profile of genotoxin-treated cells, largely mediated by the ATAXIA TELANGIECTASIA MUTATED (ATM) protein kinase, representing candidate factors that modulate plant growth, development and DNA repair. A C-terminal dual serine target motif unique to H2AX in the plant lineage showed 171-fold phosphorylation that was absent in atm mutant lines. The physiological significance of post-translational DNA damage signalling to plant growth and survival was demonstrated using reverse genetics and complementation studies of h2ax mutants, establishing the functional role of ATM-mediated histone modification in plant growth under genotoxic stress. Our findings demonstrate the complexity and functional significance of post-translational DNA damage signalling responses in plants and establish the requirement of H2AX phosphorylation for plant survival under genotoxic stress., (© 2019 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)
- Published
- 2019
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