Assembly and nuclear export of pre-ribosomal particles in budding yeast

Gerhardy, Stefan; Menet, Anna Maria; Peña, Cohue; Petkowski, Janusz J.; Panse, Vikram Govind

doi:10.1007/s00412-014-0463-z

Cited by 45 publications

(60 citation statements)

References 181 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several hundred assembly factors contribute to ribosome biogenesis by supporting the processing, modification, and folding of rRNAs, by facilitating the assembly of ribosomal proteins into pre-ribosomes, by enabling the nuclear export of pre-ribosomal particles, and by performing proofreading roles to ensure completion of upstream steps of biogenesis before later processes are initiated (Karbstein 2013;Woolford and Baserga 2013;Gerhardy et al 2014). Ribosome biogenesis starts in the nucleolus, where RNA polymerase I transcribes the 35S pre-rRNA, which contains the sequences for the 18S, 25S, and 5.8S rRNAs as well as internal and external transcribed spacers (Henras et al 2008(Henras et al , 2014.…”

Section: Introductionmentioning

confidence: 99%

“…See text for further details. Baserga 2013; Gerhardy et al 2014). While the nascent 40S subunits are rapidly exported to the cytoplasm, the biogenesis of the 60S ribosomal subunit progresses through several nucleolar and nucleoplasmic assembly and maturation steps that give rise to defined pre-ribosomal particles.…”

Section: Introductionmentioning

confidence: 99%

“…While the nascent 40S subunits are rapidly exported to the cytoplasm, the biogenesis of the 60S ribosomal subunit progresses through several nucleolar and nucleoplasmic assembly and maturation steps that give rise to defined pre-ribosomal particles. These are characterized by the arrival and departure of certain sets of assembly factors and energy-consuming enzymes as well as the accompanying structural transitions of the nascent 60S particle (Bassler et al 2001;Harnpicharnchai et al 2001;Saveanu et al 2001Saveanu et al , 2003Fatica et al 2002;Nissan et al 2002;Kressler et al 2012b;Gerhardy et al 2014). Additionally, several pre-rRNA cleavage steps, including the removal of ITS2, which is inserted between the sequences for the mature 25S and 5.8S rRNAs in the 27S pre-rRNA species, are required to generate the mature large subunit rRNAs (Henras et al 2008(Henras et al , 2014Fernandez-Pevida et al 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Upon completion of the nuclear steps of ribosome assembly, the pre-60S particle recruits nuclear export receptors and is exported to the cytoplasm, where it can enter the pool of translating ribosomes after undergoing final cytoplasmic maturation (Johnson et al 2002;Zemp and Kutay 2007;Panse and Johnson 2010). Many of the assembly factors released during cytoplasmic maturation are so-called "shuttling factors," which travel to the cytoplasm bound to the pre-60S subunit during nuclear export and return to the nucleus to participate in new rounds of ribosome biogenesis after their release from the maturing pre-ribosome (Panse and Johnson 2010;Gerhardy et al 2014).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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

Greber

2016

RNA

View full text Add to dashboard Cite

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.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Greber

2016

RNA

View full text Add to dashboard Cite

show abstract

“…The 5S rRNA is transcribed by RNA polymerase III separately (see reviews in Refs. [1][2][3][4]. In yeast, 5S, 5.8S, and 25S (28S in higher eukaryotes) rRNA assemble with 39 ribosomal proteins to make up the 60S subunit, whereas 18S rRNA and 33 ribosomal proteins constitute the 40S subunit (5-7).…”

mentioning

confidence: 99%

The Roles of Puf6 and Loc1 in 60S Biogenesis Are Interdependent, and Both Are Required for Efficient Accommodation of Rpl43

Yang

Ting

Liang

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Puf6 and Loc1 have two important functional roles in the cells, asymmetric mRNA distribution and ribosome biogenesis. Puf6 and Loc1 are localized predominantly in the nucleolus. They bind ASH1 mRNA, repress its translation, and facilitate the transport to the daughter cells. Asymmetric mRNA distribution is important for cell differentiation. Besides their roles in mRNA localization, Puf6 and Loc1 have been shown to be involved in 60S biogenesis. In puf6⌬ or loc1⌬ cells, pre-rRNA processing and 60S export are impaired and 60S subunits are underaccumulated. The functional studies of Puf6 and Loc1 have been focused on ASH1 mRNA pathway, but their roles in 60S biogenesis are still not clear. In this study, we found that Puf6 and Loc1 interact directly with each other and both proteins interact with the ribosomal protein Rpl43 (L43e). Notably, the roles of Puf6 and Loc1 in 60S biogenesis are interdependent, and both are required for efficient accommodation of Rpl43. Loc1 is further required to maintain the protein level of Rpl43. Additionally, the recruitment of Rpl43 is required for the release of Puf6 and Loc1. We propose that Puf6 and Loc1 facilitate Rpl43 loading and are sequentially released from 60S after incorporation of Rpl43 into ribosomes in yeast.Almost every process in a cell is mediated by proteins, the products of translating mRNA by ribosomes. Ribosomes are composed of two subunits, a small subunit and a large subunit, 40S and 60S, respectively, in eukaryotic cells. In Saccharomyces cerevisiae, the 40S and 60S subunits are assembled from a large primary 35 S rRNA transcribed by RNA polymerase I. Processing of this rRNA yields 5.8S, 18S, and 25S rRNAs. The 5S rRNA is transcribed by RNA polymerase III separately (see reviews in Refs. 1-4). In yeast, 5S, 5.8S, and 25S (28S in higher eukaryotes) rRNA assemble with 39 ribosomal proteins to make up the 60S subunit, whereas 18S rRNA and 33 ribosomal proteins constitute the 40S subunit (5-7).Ribosome biogenesis is necessary for cell growth and proliferation. The synthesis of ribosome is a complex pathway in which over 200 trans-acting factors are involved. Many of the RNA folding, protein assembly, and maturation events are hierarchical, requiring the correct completion of one event to progress to the next event. This is seen in the assembly of ribosomal proteins onto the RNA in bacterial ribosome assembly (8) as well as in cytoplasmic maturation of the large subunit in yeast (9, 10). In addition, trans-acting factors also perform quality control steps for the assembly of the protein synthesis machinery. Syo1 facilitates the synchronized import of the two 5S rRNA-binding proteins, Rpl5 and Rpl11, to ensure stoichiometric the incorporation of these two proteins into the pre-60S subunits and also works as an assembly platform for the 5S ribonucleoprotein (RNP) (11,12). Release of the anti-association factor Tif6 requires elongation factor-like Efl1 and tRNAlike Sdo1 to confirm the integrity of the P-site (13-15). Transacting factors on cytoplasmic pre-40S subuni...

show abstract

Eukaryotic Ribosome assembly and Nucleocytoplasmic Transport

Oborská-Oplová

Fischer

Altvater

et al. 2022

Methods in Molecular Biology

Self Cite

View full text Add to dashboard Cite

The process of eukaryotic ribosome assembly stretches across the nucleolus, the nucleoplasm and the cytoplasm, and therefore relies on efficient nucleocytoplasmic transport. In yeast, the import machinery delivers ~140,000 ribosomal proteins every minute to the nucleus for ribosome assembly. At the same time, the export machinery facilitates translocation of ~2000 pre-ribosomal particles every minute through ~200 nuclear pore complexes (NPC) into the cytoplasm. Eukaryotic ribosome assembly also requires >200 conserved assembly factors, which transiently associate with pre-ribosomal particles. Their site(s) of action on maturing pre-ribosomes are beginning to be elucidated. In this chapter, we outline protocols that enable rapid biochemical isolation of pre-ribosomal particles for single particle cryo-electron microscopy (cryo-EM) and in vitro reconstitution of nuclear transport processes. We discuss cell-biological and genetic approaches to investigate how the ribosome assembly and the nucleocytoplasmic transport machineries collaborate to produce functional ribosomes.

show abstract

Assembly and nuclear export of pre-ribosomal particles in budding yeast

Cited by 45 publications

References 181 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

The Roles of Puf6 and Loc1 in 60S Biogenesis Are Interdependent, and Both Are Required for Efficient Accommodation of Rpl43

Eukaryotic Ribosome assembly and Nucleocytoplasmic Transport

Contact Info

Product

Resources

About