title: In Situ Detection of Interactions Between Nuclear Envelope Proteins and Partners.
abstract: Proximity ligation assay (PLA) appears as a quick and easy technique to visualize within fixed cells the occurrence and in situ distribution of protein complexes. PLA has been validated to detect protein-protein interactions within the nuclear compartment. Here, we describe a protocol which allows the detection of interactions between A-type nuclear lamins and either LEM-domain proteins (such as emerin, integrated within the inner nuclear membrane, and LAP2α which accumulates within the nucleoplasm) or gene regulatory factors (e.g., the transcription factor SREBP1). The distinct amounts and patterns of PLA signals obtained for various complexes highlight the pertinence of using PLA to reveal in situ where and to which extent nuclear envelope proteins bind specific partners.
authors.affiliation: Unit of Functional and Adaptive Biology (BFA) CNRS UMR 8251, Université Paris Diderot, Sorbonne Paris Cité, 4 rue Marie-Andrée Lagroua Weill-Halle, Paris Cedex 13, 75205, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Barateau
authors.affiliation: Unit of Functional and Adaptive Biology (BFA) CNRS UMR 8251, Université Paris Diderot, Sorbonne Paris Cité, 4 rue Marie-Andrée Lagroua Weill-Halle, Paris Cedex 13, 75205, France. brigitte.buendia@univ-paris-diderot.fr.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Buendia
mesh_terms: D002352:Carrier Proteins / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D015200:Fluoroimmunoassay; D034882:Lamins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009685:Nuclear Envelope / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*
chemical_list: D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D034882:Lamins; D008565:Membrane Proteins; C093413:lamina-associated polypeptide 2
keywords: Lamins; Nuclear proteins; Proximity ligation assay