Structural and functional analysis of the Rpf2-Rrs1 complex in ribosome biogenesis

Asano, Naoki; Kato, Kuniki; Nakamura, Akiyoshi; Komoda, Keisuke; Tanaka, Isao; Yao, Min

doi:10.1093/nar/gkv305

Cited by 40 publications

(52 citation statements)

References 41 publications

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“…After binding of the 5S rRNA to the ternary Syo1-uL5-uL18 complex (Kressler et al 2012a;Calviño et al 2015), the pre-assembled 5S RNP is incorporated into very early pre-ribosomal particles aided by the assembly factor complex Rpf2-Rrs1 (Zhang et al 2007). In agreement with a role in 5S RNP recruitment and stabilization of the immature CP, the Rpf2-Rrs1 complex in the Arx1 and Nog2 pre-60S particles is located at the interface between the rotated 5S RNP and the RNA helices that later form the base of the mature CP (Asano et al 2015;Kharde et al 2015;Madru et al 2015;Wu et al 2016), along with the biogenesis factor Rsa4 (Figs. 3A, 4C,D; Leidig et al 2014).…”

Section: The Architecture Of the Immature Central Protuberance In Nucsupporting

confidence: 56%

Mechanistic insight into eukaryotic 60S ribosomal subunit biogenesis by cryo-electron microscopy

Greber

2016

RNA

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Eukaryotic ribosomes, the protein-producing factories of the cell, are composed of four ribosomal RNA molecules and roughly 80 proteins. Their biogenesis is a complex process that involves more than 200 biogenesis factors that facilitate the production, modification, and assembly of ribosomal components and the structural transitions along the maturation pathways of the preribosomal particles. Here, I review recent structural and mechanistic insights into the biogenesis of the large ribosomal subunit that were furthered by cryo-electron microscopy of natively purified pre-60S particles and in vitro reconstituted ribosome assembly factor complexes. Combined with biochemical, genetic, and previous structural data, these structures have provided detailed insights into the assembly and maturation of the central protuberance of the 60S subunit, the network of biogenesis factors near the ribosomal tunnel exit, and the functional activation of the large ribosomal subunit during cytoplasmic maturation.

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Section: The Architecture Of the Immature Central Protuberance In Nucsupporting

confidence: 56%

Mechanistic insight into eukaryotic 60S ribosomal subunit biogenesis by cryo-electron microscopy

Greber

2016

RNA

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“…In particular, main-chain tracing of the C-terminal extension of Nog1 was done completely manually. For factors Rpf2 and Rrs1, their S. cerevisiae models were generated using CHAINSAW 50 in the CCP4 suite 51 with the crystal structures of Aspergillus nidulans Rpf2 and Rrs1 (PDB accession number 4XD9 and 5BY8) 12,52 as templates. The crystal structure of the yeast Tif6 (PDB accession number 1G62) 53 was docked into the density map to provide an initial position.…”

Section: Methodsmentioning

confidence: 99%

Diverse roles of assembly factors revealed by structures of late nuclear pre-60S ribosomes

Tutuncuoglu

Yan

et al. 2016

Nature

215

499

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Ribosome biogenesis is a highly complex process in eukaryotes, involving temporally and spatially regulated ribosomal protein (r-protein) binding and ribosomal RNA remodelling events in the nucleolus, nucleoplasm and cytoplasm1,2. Hundreds of assembly factors, organized into sequential functional groups3,4, facilitate and guide the maturation process into productive assembly branches in and across different cellular compartments. However, the precise mechanisms by which these assembly factors function are largely unknown. Here we use cryo-electron microscopy to characterize the structures of yeast nucleoplasmic pre-60S particles affinity-purified using the epitope-tagged assembly factor Nog2. Our data pinpoint the locations and determine the structures of over 20 assembly factors, which are enriched in two areas: an arc region extending from the central protuberance to the polypeptide tunnel exit, and the domain including the internal transcribed spacer 2 (ITS2) that separates 5.8S and 25S ribosomal RNAs. In particular, two regulatory GTPases, Nog2 and Nog1, act as hub proteins to interact with multiple, distant assembly factors and functional ribosomal RNA elements, manifesting their critical roles in structural remodelling checkpoints and nuclear export. Moreover, our snapshots of compositionally and structurally different pre-60S intermediates provide essential mechanistic details for three major remodelling events before nuclear export: rotation of the 5S ribonucleoprotein, construction of the active centre and ITS2 removal. The rich structural information in our structures provides a framework to dissect molecular roles of diverse assembly factors in eukaryotic ribosome assembly.

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“…This structural and topological analogy defines α 1 ′ as a structural equivalent to the α 2 helix and extends the duplicated BID domain to the entire α-β-α layer. Recently, the crystal structure of the Rpf2-Rrs1 complex from Aspergillus nidulans has been reported (Asano et al 2015). Despite being from two different organisms, the structures obtained are very similar.…”

Section: Rpf2 and Rrs1 Form An Intimate Complexmentioning

confidence: 99%

Chaperoning 5S RNA assembly

Madru¹,

Lebaron²,

Blaud³

et al. 2015

Genes Dev.

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In eukaryotes, three of the four ribosomal RNAs (rRNAs)-the 5.8S, 18S, and 25S/28S rRNAs-are processed from a single pre-rRNA transcript and assembled into ribosomes. The fourth rRNA, the 5S rRNA, is transcribed by RNA polymerase III and is assembled into the 5S ribonucleoprotein particle (RNP), containing ribosomal proteins Rpl5/ uL18 and Rpl11/uL5, prior to its incorporation into preribosomes. In mammals, the 5S RNP is also a central regulator of the homeostasis of the tumor suppressor p53. The nucleolar localization of the 5S RNP and its assembly into preribosomes are performed by a specialized complex composed of Rpf2 and Rrs1 in yeast or Bxdc1 and hRrs1 in humans. Here we report the structural and functional characterization of the Rpf2-Rrs1 complex alone, in complex with the 5S RNA, and within pre-60S ribosomes. We show that the Rpf2-Rrs1 complex contains a specialized 5S RNA E-loop-binding module, contacts the Rpl5 protein, and also contacts the ribosome assembly factor Rsa4 and the 25S RNA. We propose that the Rpf2-Rrs1 complex establishes a network of interactions that guide the incorporation of the 5S RNP in preribosomes in the initial conformation prior to its rotation to form the central protuberance found in the mature large ribosomal subunit.

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Structural and functional analysis of the Rpf2-Rrs1 complex in ribosome biogenesis

Cited by 40 publications

References 41 publications

Mechanistic insight into eukaryotic 60S ribosomal subunit biogenesis by cryo-electron microscopy

Mechanistic insight into eukaryotic 60S ribosomal subunit biogenesis by cryo-electron microscopy

Diverse roles of assembly factors revealed by structures of late nuclear pre-60S ribosomes

Chaperoning 5S RNA assembly

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