Dissecting Ribosome Assembly and Transport in Budding Yeast

Altvater, Martin; Schütz, Sabina; Chang, Yeon Soo; Panse, Vikram Govind

doi:10.1016/b978-0-12-417160-2.00020-5

Cited by 16 publications

(19 citation statements)

References 42 publications

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“…Pre-40S subunit export, monitored by localization of uS5-GFP and localization of GFP-tagged Fap7 and uS11 was performed as previously described (Altvater et al, 2014; Faza et al, 2012; Pertschy et al, 2007; Milkereit et al, 2001). Cells were visualized using a DM6000B microscope (Leica, Germany) equipped with a HCX PL Fluotar 63×/1.25 NA oil immersion objective (Leica, Germany).…”

Section: Methodsmentioning

confidence: 99%

Prefabrication of a ribosomal protein subcomplex essential for eukaryotic ribosome formation

Peña

Schütz

Fischer

et al. 2016

eLife

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Spatial clustering of ribosomal proteins (r-proteins) through tertiary interactions is a striking structural feature of the eukaryotic ribosome. However, the functional importance of these intricate inter-connections, and how they are established is currently unclear. Here, we reveal that a conserved ATPase, Fap7, organizes interactions between neighboring r-proteins uS11 and eS26 prior to their delivery to the earliest ribosome precursor, the 90S. In vitro, uS11 only when bound to Fap7 becomes competent to recruit eS26 through tertiary contacts found between these r-proteins on the mature ribosome. Subsequently, Fap7 ATPase activity unloads the uS11:eS26 subcomplex onto its rRNA binding site, and therefore ensures stoichiometric integration of these r-proteins into the 90S. Fap7-depletion in vivo renders uS11 susceptible to proteolysis, and precludes eS26 incorporation into the 90S. Thus, prefabrication of a native-like r-protein subcomplex drives efficient and accurate construction of the eukaryotic ribosome.DOI: http://dx.doi.org/10.7554/eLife.21755.001

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Section: Methodsmentioning

confidence: 99%

Prefabrication of a ribosomal protein subcomplex essential for eukaryotic ribosome formation

Peña

Schütz

Fischer

et al. 2016

eLife

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“…Pre-40S subunit export, monitored by localization of uS5-GFP and localization of GFP-eS26 was performed as previously described (Faza et al, 2012; Altvater et al, 2014). Indirect immunofluorescence using affinity-purified polyclonal antibodies against the TAP-tag (1:1000; Thermo Scientific; Rockford, IL, USA) and staining of the nuclear and mitochondrial DNA with DAPI was performed as described previously (Schlenstedt et al, 1997; Solsbacher et al, 1998).…”

Section: Methodsmentioning

confidence: 99%

“…Sedimentation analysis of yeast lysates by sucrose gradient ultracentrifugation was performed as described previously (Kemmler et al, 2009; Altvater et al, 2014). For Western analyses, 500 μl fractions were precipitated by TCA (trichloroacetic acid), washed in acetone, resuspended in 100 μl of onefold SDS sample buffer and separated by SDS-PAGE.…”

Section: Methodsmentioning

confidence: 99%

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A RanGTP-independent mechanism allows ribosomal protein nuclear import for ribosome assembly

Schütz

Fischer

Altvater

et al. 2014

eLife

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114

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Within a single generation time a growing yeast cell imports ∼14 million ribosomal proteins (r-proteins) into the nucleus for ribosome production. After import, it is unclear how these intrinsically unstable and aggregation-prone proteins are targeted to the ribosome assembly site in the nucleolus. Here, we report the discovery of a conserved nuclear carrier Tsr2 that coordinates transfer of the r-protein eS26 to the earliest assembling pre-ribosome, the 90S. In vitro studies revealed that Tsr2 efficiently dissociates importin:eS26 complexes via an atypical RanGTP-independent mechanism that terminates the import process. Subsequently, Tsr2 binds the released eS26, shields it from proteolysis, and ensures its safe delivery to the 90S pre-ribosome. We anticipate similar carriers—termed here escortins—to securely connect the nuclear import machinery with pathways that deposit r-proteins onto developing pre-ribosomal particles.DOI: http://dx.doi.org/10.7554/eLife.03473.001

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“…18 The only known adaptor between Crm1 and pre-60S is Nmd3 protein 47 , while the Crm1 adaptor for pre-40S subunit has not been identified yet. 48 Mex67/Mtr2 (mRNA heterodimeric export receptor) and Arx1 proteins also participate in the export of pre-60S subunit. 49 Furthermore, other proteins have been suggested to function in the export of pre-60S subunits, e.g., the mRNA export factor Npl3 50 and Exp-5, the vertebrate orthologue of yeast Msn5 protein that is involved in tRNA transport.…”

Section: Rsp5 Affects Rrna Processing and Transportmentioning

confidence: 99%

Role of Rsp5 ubiquitin ligase in biogenesis of rRNA, mRNA and tRNA in yeast

Domanska

Kamińska

2015

RNA Biology

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Rsp5 ubiquitin ligase is required for ubiquitination of a wide variety of proteins involved in essential processes. Rsp5 was shown to be involved in regulation of lipid biosynthesis, intracellular trafficking of proteins, response to various stresses, and many other processes. In this article, we provide a comprehensive review of the nuclear and cytoplasmic functions of Rsp5 with a focus on biogenesis of different RNAs. We also briefly describe the participation of Rsp5 in the regulation of the RNA polymerase II complex, and its potential role in the regulation of other RNA polymerases. Moreover, we emphasize the function of Rsp5 in the coordination of the different steps of rRNA, mRNA and tRNA metabolism in the context of protein biosynthesis. Finally, we highlight the involvement of Rsp5 in controlling diverse cellular mechanisms at multiple levels and in adaptation of the cell to changing growth conditions.

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Dissecting Ribosome Assembly and Transport in Budding Yeast

Cited by 16 publications

References 42 publications

Prefabrication of a ribosomal protein subcomplex essential for eukaryotic ribosome formation

Prefabrication of a ribosomal protein subcomplex essential for eukaryotic ribosome formation

A RanGTP-independent mechanism allows ribosomal protein nuclear import for ribosome assembly

Role of Rsp5 ubiquitin ligase in biogenesis of rRNA, mRNA and tRNA in yeast

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