Toggle Main Menu Toggle Search

Open Access padlockePrints

Regulation of p53 and Rb Links the Alternative NF-kappa B Pathway to EZH2 Expression and Cell Senescence

Lookup NU author(s): Dr Alessio Iannetti, Dr Adeline Ledoux, Dr Susan Tudhope, Helene Sellier, Adam Moore, Dr Simon Cockell, Dr Elaine Willmore, Professor Neil Perkins

Downloads


Licence

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


Abstract

There are two major pathways leading to induction of NF-kappa B subunits. The classical (or canonical) pathway typically leads to the induction of RelA or c-Rel containing complexes, and involves the degradation of I kappa B alpha in a manner dependent on I kappa B kinase (IKK) beta and the IKK regulatory subunit NEMO. The alternative (or non-canonical) pathway, involves the inducible processing of p100 to p52, leading to the induction of NF-kappa B2(p52)/RelB containing complexes, and is dependent on IKK alpha and NF-kappa B inducing kinase (NIK). Here we demonstrate that in primary human fibroblasts, the alternative NF-kappa B pathway subunits NF-kappa B2 and RelB have multiple, but distinct, effects on the expression of key regulators of the cell cycle, reactive oxygen species (ROS) generation and protein stability. Specifically, following siRNA knockdown, quantitative PCR, western blot analyses and chromatin immunoprecipitation (ChIP) show that NF-kappa B2 regulates the expression of CDK4 and CDK6, while RelB, through the regulation of genes such as PSMA5 and ANAPC1, regulates the stability of p21WAF1 and the tumour suppressor p53. These combine to regulate the activity of the retinoblastoma protein, Rb, leading to induction of polycomb protein EZH2 expression. Moreover, our ChIP analysis demonstrates that EZH2 is also a direct NF-kappa B target gene. Microarray analysis revealed that in fibroblasts, EZH2 antagonizes a subset of p53 target genes previously associated with the senescent cell phenotype, including DEK and RacGAP1. We show that this pathway provides the major route of crosstalk between the alternative NF-kappa B pathway and p53, a consequence of which is to suppress cell senescence. Importantly, we find that activation of NF-kappa B also induces EZH2 expression in CD40L stimulated cells from Chronic Lymphocytic Leukemia patients. We therefore propose that this pathway provides a mechanism through which microenvironment induced NF-kappa B can inhibit tumor suppressor function and promote tumorigenesis.


Publication metadata

Author(s): Iannetti A, Ledoux AC, Tudhope SJ, Sellier H, Zhao B, Mowla S, Moore A, Hummerich H, Gewurz BE, Cockell SJ, Jat PS, Willmore E, Perkins ND

Publication type: Article

Publication status: Published

Journal: PLoS Genetics

Year: 2014

Volume: 10

Issue: 9

Print publication date: 01/09/2014

Online publication date: 25/09/2014

Acceptance date: 28/07/2014

Date deposited: 08/12/2014

ISSN (print): 1553-7390

ISSN (electronic): 1553-7404

Publisher: Public Library of Science

URL: http://dx.doi.org/10.1371/journal.pgen.1004642

DOI: 10.1371/journal.pgen.1004642


Altmetrics

Altmetrics provided by Altmetric


Actions

Find at Newcastle University icon    Link to this publication


Share