Chaperoning 5S RNA assembly

Madru, Clément; Lebaron, Simon; Blaud, Magali; Delbos, Lila; Pipoli, J.; Pasmant, Éric; Réty, S.; Leulliot, Nicolas

doi:10.1101/gad.260349.115

Cited by 57 publications

(80 citation statements)

References 53 publications

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“…Interestingly, Rpf2, Rrs1, and Rsa4 form a network that drives formation of the final structure of H89 in rRNA domain V (Kharde et al 2015;Madru et al 2015, Wu et al 2016. Since all of these assembly factors interact with rRNA domain V in the assembling 60S subunit, our results further confirm the role of the N-terminal extension of L8 in structuring domain V (Fig.…”

Section: Discussionsupporting

confidence: 76%

The N-terminal extension of yeast ribosomal protein L8 is involved in two major remodeling events during late nuclear stages of 60S ribosomal subunit assembly

Tutuncuoglu

Jakovljevic

et al. 2016

RNA

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Assaying effects on pre-rRNA processing and ribosome assembly upon depleting individual ribosomal proteins (r-proteins) provided an initial paradigm for assembly of eukaryotic ribosomes in vivo-that each structural domain of ribosomal subunits assembles in a hierarchical fashion. However, two features suggest that a more complex pathway may exist: (i) Some r-proteins contain extensions that reach long distances across ribosomes to interact with multiple rRNA domains as well as with other r-proteins. (ii) Individual r-proteins may assemble in a stepwise fashion. For example, the globular domain of an r-protein might assemble separately from its extensions. Thus, these extensions might play roles in assembly that could not be revealed by depleting the entire protein. Here, we show that deleting or mutating extensions of r-proteins L7 (uL30) and L35 (uL29) from yeast reveal important roles in early and middle steps during 60S ribosomal subunit biogenesis. Detailed analysis of the N-terminal terminal extension of L8 (eL8) showed that it is necessary for late nuclear stages of 60S subunit assembly involving two major remodeling events: removal of the ITS2 spacer; and reorganization of the central protuberance (CP) containing 5S rRNA and r-proteins L5 (uL18) and L11 (uL5). Mutations in the L8 extension block processing of 7S pre-rRNA, prevent release of assembly factors Rpf2 and Rrs1 from pre-ribosomes, which is required for rotation of the CP, and block association of Sda1, the Rix1 complex, and the Rea1 ATPase involved in late steps of remodeling.

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Section: Discussionsupporting

confidence: 76%

The N-terminal extension of yeast ribosomal protein L8 is involved in two major remodeling events during late nuclear stages of 60S ribosomal subunit assembly

Tutuncuoglu

Jakovljevic

et al. 2016

RNA

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“…4D) and confirmed that Rrs1 interacts with the protein components of the 5S RNP and the biogenesis factor Rsa4, while Rpf2 forms extensive interactions with the 5S rRNA, uL18, and Rsa4 ( Fig. 4C,D; Kharde et al 2015;Madru et al 2015;Wu et al 2016).…”

Section: The Architecture Of the Immature Central Protuberance In Nucmentioning

confidence: 59%

“…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 Nucmentioning

confidence: 74%

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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“…Formation of the 5S RNP is facilitated by the transport adaptor Syo1, which couples nuclear import of uL5 and uL18 with 5S rRNA association [90,91]. The subsequent incorporation of the 5S RNP module involves the Rpf2-Rrs1 heterodimer and has already occurred within the Nsa1-defined particle [25,86,[92][93][94] (Figure 2K). However, the exact timing and the mechanistic details of the initial 5S RNP docking to the pre-60S ribosome remain to be determined.…”

Section: Assembly Of the Lsumentioning

confidence: 99%

A Puzzle of Life: Crafting Ribosomal Subunits

Kressler

Hurt

Baßler

2017

Trends in Biochemical Sciences

164

127

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The biogenesis of eukaryotic ribosomes is a complicated process during which the transcription, modification, folding, and processing of the rRNA is coupled with the ordered assembly of $80 ribosomal proteins (r-proteins). Ribosome synthesis is catalyzed and coordinated by more than 200 biogenesis factors as the preribosomal subunits acquire maturity on their path from the nucleolus to the cytoplasm. Several biogenesis factors also interconnect the progression of ribosome assembly with quality control of important domains, ensuring that only functional subunits engage in translation. With the recent visualization of several assembly intermediates by cryoelectron microscopy (cryo-EM), a structural view of ribosome assembly begins to emerge. In this review we integrate these first structural insights into an updated overview of the consecutive ribosome assembly steps. Synopsis of Eukaryotic Ribosome AssemblyRibosomes are the molecular machines that translate the genetic information from the intermediary mRNA templates into proteins [1]. Eukaryotic 80S ribosomes comprise two unequal subunits that contain four different rRNAs and around 80 r-proteins ( Figure 1). The small 40S subunit (SSU) comprises the 18S rRNA and 33 r-proteins (referred to as RPS or S). The large 60S subunit (LSU) comprises the 25S/28S, 5.8S, and 5S rRNA and, in most eukaryotic species, 47 r-proteins (RPL or L); a notable exception is budding yeasts, which lack eL28The act of building a ribosome begins in the nucleolus, where the ribosomal DNA (rDNA) is transcribed into a long pre-rRNA precursor (35S pre-rRNA in yeast) and involves the ordered assembly of the r-proteins with the pre-rRNA, which is concomitantly processed into the mature rRNA species [5][6][7][8] (Figure 1 and Box 1). These assembly and processing events are tightly coupled and occur within preribosomal particles (see Glossary) that travel, as maturation progresses, from the nucleus across nuclear pore complexes (NPCs) to the cytoplasm, where they are ultimately converted into translation-competent ribosomal subunits [5,[9][10][11][12][13] (Figure 2, Key Figure). Given the gargantuan complexity of this process, it is unsurprising that the assembly of eukaryotic ribosomes strictly requires the assistance of a plethora (>200) of mostly essential ribosome biogenesis factors, which are also called trans-acting or assembly factors [5,14,15].Our current knowledge has been mainly obtained by studying ribosome biogenesis in the yeast Saccharomyces cerevisiae. Research conducted in the 1970s defined the r-protein composition of ribosomes, revealed the major pre-rRNA processing intermediates, and uncovered the existence of the 90S, 43S, and 66S preribosomal particles. The following 25 years witnessed the identification of numerous biogenesis factors and attributed functional roles TrendsCryoelectron microscopy analyses of the 90S preribosome show that the biogenesis factors create a casting mold that encloses the nascent pre-40S subunit.Structures of nuclear pre-60S particles reveal that a...

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Chaperoning 5S RNA assembly

Cited by 57 publications

References 53 publications

The N-terminal extension of yeast ribosomal protein L8 is involved in two major remodeling events during late nuclear stages of 60S ribosomal subunit assembly

The N-terminal extension of yeast ribosomal protein L8 is involved in two major remodeling events during late nuclear stages of 60S ribosomal subunit assembly

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

A Puzzle of Life: Crafting Ribosomal Subunits

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