1. Drosophila Spag Is the Homolog of RNA Polymerase II-associated Protein 3 (RPAP3) and Recruits the Heat Shock Proteins 70 and 90 (Hsp70 and Hsp90) during the Assembly of Cellular Machineries
- Author
-
Ioannis Iliopoulos, Nour El Houda Benbahouche, Tore Kempf, Istvan Török, Philippe Meyer, Robert Farkaš, Edouard Bertrand, István Kiss, Julien Henri, Bernard M. Mechler, Bérengère Pradet-Balade, Andrey V. Kajava, Joachim Marhold, Martina Schnölzer, Institut d'Électronique et des Technologies du numéRique (IETR), Université de Nantes (UN)-Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Computationnelle (IBC), Institut National de la Recherche Agronomique (INRA)-Institut National de Recherche en Informatique et en Automatique (Inria)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche en Biologie Cellulaire (CRBM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), National Research University of Information Technologies, Mechanics and Optics [St. Petersburg] (ITMO), STMicroelectronics [Rousset] (ST-ROUSSET), Institut des Matériaux, de Microélectronique et des Nanosciences de Provence (IM2NP), Aix Marseille Université (AMU)-Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Polytech Nice-Sophia (Polytech'Lab), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), CRLCC Paul Strauss, Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Nantes (UN)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS), Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1), Université de Toulon (UTLN)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Université Nice Sophia Antipolis (... - 2019) (UNS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut d'Electronique et de Télécommunications de Rennes (IETR), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre de recherches de biochimie macromoléculaire (CRBM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-IFR122-Centre National de la Recherche Scientifique (CNRS), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA), and Nantes Université (NU)-Université de Rennes 1 (UR1)
- Subjects
Animals Anti-Bacterial Agents/pharmacology Carrier Proteins/genetics/metabolism Drosophila Proteins/genetics/*metabolism Drosophila melanogaster HSP70 Heat-Shock Proteins/genetics/*metabolism Heat-Shock Proteins/genetics/*metabolism Humans *Models ,Saccharomyces cerevisiae Proteins ,RNA polymerase II ,Saccharomyces cerevisiae ,Biology ,Models, Biological ,Biochemistry ,03 medical and health sciences ,Ribonucleoproteins, Small Nucleolar ,RNA interference ,RUVBL2 ,Animals ,Drosophila Proteins ,Humans ,HSP70 Heat-Shock Proteins ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Small nucleolar RNA ,Molecular Biology ,Heat-Shock Proteins ,ComputingMilieux_MISCELLANEOUS ,Polymerase ,030304 developmental biology ,R2TP complex ,Sirolimus ,Genetics ,0303 health sciences ,030302 biochemistry & molecular biology ,RNA ,Cell Biology ,Small Nucleolar/genetics/*metabolism Saccharomyces cerevisiae/genetics/metabolism Saccharomyces cerevisiae Proteins/genetics/metabolism Sirolimus/pharmacology Hsp70 Hsp90 Protein Assembly R2TP Complex RNA Polymerase II Rpap3 Small Nucleolar RNA (snoRNA) Spaghetti Gene TOR Complex (TORC) ,Anti-Bacterial Agents ,Cell biology ,Drosophila melanogaster ,Protein Synthesis and Degradation ,biology.protein ,Biological Molecular Chaperones/genetics/*metabolism RNA Polymerase II/genetics/metabolism Ribonucleoproteins ,RNA Polymerase II ,Apoptosis Regulatory Proteins ,Carrier Proteins ,Drosophila Protein ,Molecular Chaperones - Abstract
The R2TP is a recently identified Hsp90 co-chaperone, composed of four proteins as follows: Pih1D1, RPAP3, and the AAA(+)-ATPases RUVBL1 and RUVBL2. In mammals, the R2TP is involved in the biogenesis of cellular machineries such as RNA polymerases, small nucleolar ribonucleoparticles and phosphatidylinositol 3-kinase-related kinases. Here, we characterize the spaghetti (spag) gene of Drosophila, the homolog of human RPAP3. This gene plays an essential function during Drosophila development. We show that Spag protein binds Drosophila orthologs of R2TP components and Hsp90, like its yeast counterpart. Unexpectedly, Spag also interacts and stimulates the chaperone activity of Hsp70. Using null mutants and flies with inducible RNAi, we show that spaghetti is necessary for the stabilization of snoRNP core proteins and target of rapamycin activity and likely the assembly of RNA polymerase II. This work highlights the strong conservation of both the HSP90/R2TP system and its clients and further shows that Spag, unlike Saccharomyces cerevisiae Tah1, performs essential functions in metazoans. Interaction of Spag with both Hsp70 and Hsp90 suggests a model whereby R2TP would accompany clients from Hsp70 to Hsp90 to facilitate their assembly into macromolecular complexes.
- Published
- 2014