title: A network of assembly factors is involved in remodeling rRNA elements during preribosome maturation.
abstract: Eukaryotic ribosome biogenesis involves ∼200 assembly factors, but how these contribute to ribosome maturation is poorly understood. Here, we identify a network of factors on the nascent 60S subunit that actively remodels preribosome structure. At its hub is Rsa4, a direct substrate of the force-generating ATPase Rea1. We show that Rsa4 is connected to the central protuberance by binding to Rpl5 and to ribosomal RNA (rRNA) helix 89 of the nascent peptidyl transferase center (PTC) through Nsa2. Importantly, Nsa2 binds to helix 89 before relocation of helix 89 to the PTC. Structure-based mutations of these factors reveal the functional importance of their interactions for ribosome assembly. Thus, Rsa4 is held tightly in the preribosome and can serve as a "distribution box," transmitting remodeling energy from Rea1 into the developing ribosome. We suggest that a relay-like factor network coupled to a mechano-enzyme is strategically positioned to relocate rRNA elements during ribosome maturation.
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Baßler
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Helge
authors.initials: H
authors.lastname: Paternoga
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Iris
authors.initials: I
authors.lastname: Holdermann
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Thoms
authors.affiliation: Centre for Synthetic and Systems Biology (SynthSys) and Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, Scotland, UK.
authors.forename: Sander
authors.initials: S
authors.lastname: Granneman
authors.affiliation: Gene Center, Department of Chemistry and Biochemistry, University of Munich, 80539 Munich, Germany.
authors.forename: Clara
authors.initials: C
authors.lastname: Barrio-Garcia
authors.affiliation: Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331.
authors.forename: Afua
authors.initials: A
authors.lastname: Nyarko
authors.affiliation: National Magnetic Resonance Facility; Biochemistry Department, University of Wisconsin-Madison, Madison, WI 53706.
authors.forename: Woonghee
authors.initials: W
authors.lastname: Lee
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Gunter
authors.initials: G
authors.lastname: Stier
authors.affiliation: Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Clark
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Schraivogel
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany.
authors.forename: Martina
authors.initials: M
authors.lastname: Kallas
authors.affiliation: Gene Center, Department of Chemistry and Biochemistry, University of Munich, 80539 Munich, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Beckmann
authors.affiliation: Centre for Synthetic and Systems Biology (SynthSys) and Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, Scotland, UK.
authors.forename: David
authors.initials: D
authors.lastname: Tollervey
authors.affiliation: Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331 ed.hurt@bzh.uni-heidelberg.de irmi.sinning@bzh.uni-heidelberg.de barbare@science.oregonstate.edu.
authors.forename: Elisar
authors.initials: E
authors.lastname: Barbar
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany ed.hurt@bzh.uni-heidelberg.de irmi.sinning@bzh.uni-heidelberg.de barbare@science.oregonstate.edu.
authors.forename: Irmi
authors.initials: I
authors.lastname: Sinning
authors.affiliation: Biochemistry Center of Heidelberg University, INF328, D-69120 Heidelberg, Germany ed.hurt@bzh.uni-heidelberg.de irmi.sinning@bzh.uni-heidelberg.de barbare@science.oregonstate.edu.
authors.forename: Ed
authors.initials: E
authors.lastname: Hurt
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000235:genetics; D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D012335:RNA, Ribosomal / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics* / Q000648:ultrastructure; D054683:Ribosome Subunits, Large, Eukaryotic / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000648:ultrastructure; D016415:Sequence Alignment
chemical_list: C514434:Nsa2 protein, S cerevisiae; D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; C489268:RPL5 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; C504806:Rsa4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C031127:ribosomal protein L25; D000251:Adenosine Triphosphatases; C496538:MDN1 protein, S cerevisiae; D000074183:ATPases Associated with Diverse Cellular Activities