caa a22 4500 606175083 CHVBK 20210128100741.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00018-014-1745-6 doi (NATIONALLICENCE)springer-10.1007/s00018-014-1745-6 ARTC1-mediated ADP-ribosylation of GRP78/BiP: a new player in endoplasmic-reticulum stress responses [Elektronische Daten] [Gaia Fabrizio, Simone Di Paola, Annalisa Stilla, Monica Giannotta, Carmen Ruggiero, Stephan Menzel, Friedrich Koch-Nolte, Michele Sallese, Maria Di Girolamo] Protein mono-ADP-ribosylation is a reversible post-translational modification of cellular proteins. This scheme of amino-acid modification is used not only by bacterial toxins to attack host cells, but also by endogenous ADP-ribosyltransferases (ARTs) in mammalian cells. These latter ARTs include members of three different families of proteins: the well characterised arginine-specific ecto-enzymes (ARTCs), two sirtuins, and some members of the poly(ADP-ribose) polymerase (PARP/ARTD) family. In the present study, we demonstrate that human ARTC1 is localised to the endoplasmic reticulum (ER), in contrast to the previously characterised ARTC proteins, which are typical GPI-anchored ecto-enzymes. Moreover, using the "macro domain” cognitive binding module to identify ADP-ribosylated proteins, we show here that the ER luminal chaperone GRP78/BiP (glucose-regulated protein of 78kDa/immunoglobulin heavy-chain-binding protein) is a cellular target of human ARTC1 and hamster ARTC2. We further developed a procedure to visualise ADP-ribosylated proteins using immunofluorescence. With this approach, in cells overexpressing ARTC1, we detected staining of the ER that co-localises with GRP78/BiP, thus confirming that this modification occurs in living cells. In line with the key role of GRP78/BiP in the ER stress response system, we provide evidence here that ARTC1 is activated during the ER stress response, which results in acute ADP-ribosylation of GRP78/BiP paralleling translational inhibition. Thus, this identification of ARTC1 as a regulator of GRP78/BiP defines a novel, previously unsuspected, player in GRP78-mediated ER stress responses. Springer Basel, 2014 ADP-ribosyltransferase nationallicence Post-translational modification nationallicence ADP-ribosylation nationallicence Macro domain nationallicence Endoplasmic reticulum nationallicence GRP78/BiP nationallicence Dithiothreitol nationallicence Thapsigargin nationallicence Chaperone nationallicence ART : ADP-ribosyltransferase nationallicence CHO cells : Chinese hamster ovary cells nationallicence DTT : Dithiothreitol nationallicence ER : Endoplasmic reticulum nationallicence GST : Glutatione S-transferase nationallicence HeLa cells : Human cervix adenocarcinoma cells nationallicence Fabrizio Gaia Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Di Paola Simone Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Stilla Annalisa Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Giannotta Monica Genomic Approaches to Membrane Traffic Unit, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale, 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Ruggiero Carmen Genomic Approaches to Membrane Traffic Unit, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale, 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Menzel Stephan Institute of Immunology, University Medical Centre Hamburg-Eppendorf, Martinist 52, 20246, Hamburg, Germany aut Koch-Nolte Friedrich Institute of Immunology, University Medical Centre Hamburg-Eppendorf, Martinist 52, 20246, Hamburg, Germany aut Sallese Michele Genomic Approaches to Membrane Traffic Unit, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale, 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Di Girolamo Maria Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut Cellular and Molecular Life Sciences Springer Basel 72/6(2015-03-01), 1209-1225 1420-682X 72:6<1209 2015 72 18 https://doi.org/10.1007/s00018-014-1745-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-014-1745-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Fabrizio Gaia Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 700 1- Di Paola Simone Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 700 1- Stilla Annalisa Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 700 1- Giannotta Monica Genomic Approaches to Membrane Traffic Unit, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale, 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 700 1- Ruggiero Carmen Genomic Approaches to Membrane Traffic Unit, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale, 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 700 1- Menzel Stephan Institute of Immunology, University Medical Centre Hamburg-Eppendorf, Martinist 52, 20246, Hamburg, Germany aut NATIONALLICENCE 700 1- Koch-Nolte Friedrich Institute of Immunology, University Medical Centre Hamburg-Eppendorf, Martinist 52, 20246, Hamburg, Germany aut NATIONALLICENCE 700 1- Sallese Michele Genomic Approaches to Membrane Traffic Unit, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale, 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 700 1- Di Girolamo Maria Laboratory of G-Protein-mediated Signalling, Department of Cellular and Translational Pharmacology, Mario Negri Sud Foundation, Via Nazionale 8/A, 66030, Santa Maria Imbaro, CH, Italy aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/6(2015-03-01), 1209-1225 1420-682X 72:6<1209 2015 72 18