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Lookup NU author(s): Dr Ling-I Su, Emma Corbin, Professor Neil PerkinsORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2021 Portland Press Ltd. All rights reserved.Different types of DNA damage can initiate phosphorylation-mediated signalling cascades that result in stimulus specific pro-or anti-Apoptotic cellular responses. Amongst its many roles, the NF-B transcription factor RelA is central to these DNA damage response pathways. However, we still lack understanding of the co-ordinated signalling mechanisms that permit different DNA damaging agents to induce distinct cellular outcomes through RelA. Here, we use label-free quantitative phosphoproteomics to examine the temporal effects of exposure of U2OS cells to either etoposide (ETO) or hydroxyurea (HU) by monitoring the phosphorylation status of RelA and its protein binding partners. Although few stimulus-specific differences were identified in the constituents of phosphorylated RelA interactome after exposure to these DNA damaging agents, we observed subtle, but significant, changes in their phosphorylation states, as a function of both type and duration of treatment. The DNA double strand break (DSB)-inducing ETO invoked more rapid, sustained responses than HU, with regulated targets primarily involved in transcription, cell division and canonical DSB repair. Kinase substrate prediction of ETOregulated phosphosites suggest abrogation of CDK and ERK1 signalling, in addition to the known induction of ATM/ATR. In contrast, HU-induced replicative stress mediated temporally dynamic regulation, with phosphorylated RelA binding partners having roles in rRNA/mRNA processing and translational initiation, many of which contained a 14-3-3 binding motif, and were putative substrates of the dual specificity kinase CLK1. Our data thus point to differential regulation of key cellular processes and the involvement of distinct signalling pathways in modulating DNA damage-specific functions of RelA.
Author(s): Campbell AE, Franco CF, Su L-I, Corbin EK, Perkins S, Kalyuzhnyy A, Jones AR, Brownridge PJ, Perkins ND, Eyers CE
Publication type: Article
Publication status: Published
Journal: Biochemical Journal
Year: 2021
Volume: 478
Issue: 3
Pages: 533-551
Print publication date: 01/02/2021
Online publication date: 13/01/2021
Acceptance date: 12/01/2021
Date deposited: 30/09/2022
ISSN (print): 0264-6021
ISSN (electronic): 1470-8728
Publisher: Portland Press Ltd
URL: https://doi.org/10.1042/BCJ20200627
DOI: 10.1042/BCJ20200627
PubMed id: 33438746
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