title: Regulation of p53 and Rb links the alternative NF-κB pathway to EZH2 expression and cell senescence.
abstract: There are two major pathways leading to induction of NF-κB subunits. The classical (or canonical) pathway typically leads to the induction of RelA or c-Rel containing complexes, and involves the degradation of IκBα in a manner dependent on IκB kinase (IKK) β 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-κB2(p52)/RelB containing complexes, and is dependent on IKKα and NF-κB inducing kinase (NIK). Here we demonstrate that in primary human fibroblasts, the alternative NF-κB pathway subunits NF-κB2 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-κB2 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-κB 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-κB pathway and p53, a consequence of which is to suppress cell senescence. Importantly, we find that activation of NF-κB 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-κB can inhibit tumor suppressor function and promote tumorigenesis.
authors.affiliation: Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Alessio
authors.initials: A
authors.lastname: Iannetti
authors.affiliation: Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Adeline C
authors.initials: AC
authors.lastname: Ledoux
authors.affiliation: Northern Institute for Cancer Research, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Tudhope
authors.affiliation: Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Hélène
authors.initials: H
authors.lastname: Sellier
authors.affiliation: Division of Infectious Disease, Brigham and Women's Hospital, Boston, Massachusetts, United States of America.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhao
authors.affiliation: Department of Neurodegenerative Disease, UCL Institute of Neurology, London, United Kingdom.
authors.forename: Sophia
authors.initials: S
authors.lastname: Mowla
authors.affiliation: Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Adam
authors.initials: A
authors.lastname: Moore
authors.affiliation: Department of Neurodegenerative Disease, UCL Institute of Neurology, London, United Kingdom.
authors.forename: Holger
authors.initials: H
authors.lastname: Hummerich
authors.affiliation: Division of Infectious Disease, Brigham and Women's Hospital, Boston, Massachusetts, United States of America.
authors.forename: Benjamin E
authors.initials: BE
authors.lastname: Gewurz
authors.affiliation: Bioinformatics Support Unit, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Cockell
authors.affiliation: Department of Neurodegenerative Disease, UCL Institute of Neurology, London, United Kingdom.
authors.forename: Parmjit S
authors.initials: PS
authors.lastname: Jat
authors.affiliation: Northern Institute for Cancer Research, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Elaine
authors.initials: E
authors.lastname: Willmore
authors.affiliation: Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle Upon Tyne, United Kingdom.
authors.forename: Neil D
authors.initials: ND
authors.lastname: Perkins
mesh_terms: D023201:CD40 Ligand / Q000819:agonists / Q000378:metabolism; D016922:Cellular Senescence / Q000235:genetics*; D016000:Cluster Analysis; D000071221:Enhancer of Zeste Homolog 2 Protein; D004789:Enzyme Activation; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D016328:NF-kappa B / Q000378:metabolism*; D052003:NF-kappa B p52 Subunit / Q000378:metabolism; D063151:Polycomb Repressive Complex 2 / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D055550:Protein Stability; D034622:RNA Interference; D017382:Reactive Oxygen Species / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism*; D015398:Signal Transduction; D051841:Transcription Factor RelB / Q000378:metabolism; D014158:Transcription, Genetic; D059467:Transcriptome; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D016328:NF-kappa B; D052003:NF-kappa B p52 Subunit; D017382:Reactive Oxygen Species; D016160:Retinoblastoma Protein; D016159:Tumor Suppressor Protein p53; D023201:CD40 Ligand; D051841:Transcription Factor RelB; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2