title: Structural analysis of the yeast exosome Rrp6p-Rrp47p complex by small-angle X-ray scattering.
abstract: The RNase D-type 3'-5' exonuclease Rrp6p from Saccharomyces cerevisiae is a nuclear-specific cofactor of the RNA exosome and associates in vivo with Rrp47p (Lrp1p). Here, we show using biochemistry and small-angle X-ray scattering (SAXS) that Rrp6p and Rrp47p associate into a stable, heterodimeric complex with an elongated shape consistent with binding of Rrp47p to the nuclease domain and opposite of the HRDC domain of Rrp6p. Rrp47p reduces the exonucleolytic activity of Rrp6p on both single-stranded and structured RNA substrates without significantly altering the affinity towards RNA or the ability of Rrp6p to degrade RNA secondary structure.
authors.affiliation: Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Emil
authors.initials: E
authors.lastname: Dedic
authors.affiliation: Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Paulina
authors.initials: P
authors.lastname: Seweryn
authors.affiliation: Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Anette Thyssen
authors.initials: AT
authors.lastname: Jonstrup
authors.affiliation: Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Rasmus Koch
authors.initials: RK
authors.lastname: Flygaard
authors.affiliation: Department of Biomedicine, Ole Worms Allé 6, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Natalya U
authors.initials: NU
authors.lastname: Fedosova
authors.affiliation: Institute for Storage Ring Facilities (ISA), Department of Physics and Astronomy, Ny Munkegade 120, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Søren Vrønning
authors.initials: SV
authors.lastname: Hoffmann
authors.affiliation: Centre for Membrane Pumps in Cells and Disease - PUMPKIN, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark.
authors.forename: Thomas
authors.initials: T
authors.lastname: Boesen
authors.affiliation: Centre for mRNP Biogenesis and Metabolism, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Gustav Wieds Vej 10c, Aarhus University, DK-8000 Aarhus C, Denmark. Electronic address: deb@mb.au.dk.
authors.forename: Ditlev Egeskov
authors.initials: DE
authors.lastname: Brodersen
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003198:Computer Simulation; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D063326:Exosome Multienzyme Ribonuclease Complex / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D011485:Protein Binding; D011487:Protein Conformation; D012313:RNA / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
chemical_list: D004268:DNA-Binding Proteins; C494789:LRP1 protein, S cerevisiae; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D012313:RNA; D063326:Exosome Multienzyme Ribonuclease Complex; C112610:RRP6 protein, S cerevisiae
keywords: 3′–5′ nuclease; Exosome; RNA processing; RNA turnover; Ribosomal RNA; snoRNA