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Dynamic phosphorylation of RelA on Ser42 and Ser45 in response to TNFα stimulation regulates DNA binding and transcription

Lookup NU author(s): Professor Jelena Mann, Professor Derek Mann

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

The NF-kappa B signalling module controls transcription through a network of protein kinases such as the IKKs, aswell as inhibitory proteins (I kappa Bs) and transcription factors including RelA/p65. Phosphorylation of the NF-kappa B subunits is critical for dictating system dynamics. Using both non-targeted discovery and quantitative selected reaction monitoring-targeted proteomics, we show that the cytokine TNF alpha induces dynamic multisite phosphorylation of RelA at a number of previously unidentified residues. Putative roles for many of these phosphorylation sites on RelA were predicted by modelling of various crystal structures. Stoichiometry of phosphorylation determination of Ser45 and Ser42 revealed preferential early phosphorylation of Ser45 in response to TNF alpha. Quantitative analyses subsequently confirmed differential roles for pSer42 and pSer45 in promoter-specific DNA binding and a role for both of these phosphosites in regulating transcription from the IL-6 promoter. These temporal dynamics suggest that RelA-mediated transcription is likely to be controlled by functionally distinct NF-kappa B proteoforms carrying different combinations of modifications, rather than a simple 'one modification, one effect' system.


Publication metadata

Author(s): Lanucara F, Lam C, Mann J, Monie TP, Colombo SAP, Holman SW, Boyd J, Dange MC, Mann DA, White MRH, Eyers CE

Publication type: Article

Publication status: Published

Journal: Open Biology

Year: 2016

Volume: 6

Issue: 7

Online publication date: 27/07/2016

Acceptance date: 01/07/2016

ISSN (electronic): 2046-2441

Publisher: The Royal Society Publishing

URL: http://dx.doi.org/10.1098/rsob.160055

DOI: 10.1098/rsob.160055

PubMed id: 27466442


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