title: Super-resolution microscopy reveals a preformed NEMO lattice structure that is collapsed in incontinentia pigmenti.
abstract: The NF-κB pathway has critical roles in cancer, immunity and inflammatory responses. Understanding the mechanism(s) by which mutations in genes involved in the pathway cause disease has provided valuable insight into its regulation, yet many aspects remain unexplained. Several lines of evidence have led to the hypothesis that the regulatory/sensor protein NEMO acts as a biological binary switch. This hypothesis depends on the formation of a higher-order structure, which has yet to be identified using traditional molecular techniques. Here we use super-resolution microscopy to reveal the existence of higher-order NEMO lattice structures dependent on the presence of polyubiquitin chains before NF-κB activation. Such structures may permit proximity-based trans-autophosphorylation, leading to cooperative activation of the signalling cascade. We further show that NF-κB activation results in modification of these structures. Finally, we demonstrate that these structures are abrogated in cells derived from incontinentia pigmenti patients.
authors: Faculty of Health Sciences, Division of Chemical Systems and Synthetic Biology, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, Western Cape 7925, South Africa.|Gene Expression and Biophysics Group, CSIR Synthetic Biology ERA, Pretoria 0001, South Africa.
authors: Janine
authors: J
authors: Scholefield
authors: Quantitative Imaging and Nanobiophysics Group, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors: Ricardo
authors: R
authors: Henriques
authors: Faculty of Health Sciences, Division of Chemical Systems and Synthetic Biology, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, Western Cape 7925, South Africa.|Gene Expression and Biophysics Group, CSIR Synthetic Biology ERA, Pretoria 0001, South Africa.
authors: Anca F
authors: AF
authors: Savulescu
authors: Chemogenomic and Biological Screening Core Facility, Institut Pasteur, Center for Innovation and Technological Research (Citech), Departments of Cell Biology and Infection and of Structural Biology and Chemistry, 25/28 rue du Dr Roux, 75724 Paris cedex 15, France.
authors: Elisabeth
authors: E
authors: Fontan
authors: Chemogenomic and Biological Screening Core Facility, Institut Pasteur, Center for Innovation and Technological Research (Citech), Departments of Cell Biology and Infection and of Structural Biology and Chemistry, 25/28 rue du Dr Roux, 75724 Paris cedex 15, France.
authors: Alix
authors: A
authors: Boucharlat
authors: Laboratory of Signaling and Pathogenesis, CNRS, UMR 3691, Institut Pasteur, 75724 Paris cedex 15, France.
authors: Emmanuel
authors: E
authors: Laplantine
authors: INSERM U1163-Université Paris Descartes-Sorbonne Paris Cité, Institut Imagine, Hôpital Necker-Enfants Malades, 24 Boulevard du Montparnasse, Paris 75015, France.
authors: Asma
authors: A
authors: Smahi
authors: Laboratory of Signaling and Pathogenesis, CNRS, UMR 3691, Institut Pasteur, 75724 Paris cedex 15, France.
authors: Alain
authors: A
authors: Israël
authors: Chemogenomic and Biological Screening Core Facility, Institut Pasteur, Center for Innovation and Technological Research (Citech), Departments of Cell Biology and Infection and of Structural Biology and Chemistry, 25/28 rue du Dr Roux, 75724 Paris cedex 15, France.
authors: Fabrice
authors: F
authors: Agou
authors: Faculty of Health Sciences, Division of Chemical Systems and Synthetic Biology, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Anzio Road, Observatory, Cape Town, Western Cape 7925, South Africa.|Gene Expression and Biophysics Group, CSIR Synthetic Biology ERA, Pretoria 0001, South Africa.|Faculdade de Medicina, Gene Expression and Biophysics Unit, Instituto de Medicina Molecular, Universidade de Lisboa, Lisbon 1649-028, Portugal.
authors: Musa M
authors: MM
authors: Mhlanga
mesh_terms: D045744:Cell Line, Tumor; D004789:Enzyme Activation; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure*; D007184:Incontinentia Pigmenti / Q000473:pathology*; D008853:Microscopy / Q000379:methods*; D016328:NF-kappa B / Q000378:metabolism*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D025801:Ubiquitin / Q000378:metabolism
chemical_list: C491731:IKBKG protein, human; D016328:NF-kappa B; D025801:Ubiquitin; D051550:I-kappa B Kinase