Nuclear import of dimerized ribosomal protein Rps3 in complex with its chaperone Yar1

Mitterer, Valentin; Gantenbein, Nadine; Birner‐Gruenberger, Ruth; Murat, Guillaume; Bergler, Helmut; Kressler, Dieter; Pertschy, Brigitte

doi:10.1038/srep36714

Cited by 28 publications

(28 citation statements)

References 46 publications

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“…As shown in Fig. 2B, uS5 was efficiently recovered in anti-GFP precipitates prepared from extracts of cells that expressed the WT version of ZNF277 (see lane 9). In contrast, a control purification prepared from extracts of cells that expressed GFP alone did not copurify uS5 (Fig.…”

Section: C2h2 Zinc Finger Domains Of Znf277 Are Important For the Assmentioning

confidence: 80%

The 40S ribosomal protein uS5 (RPS2) assembles into an extraribosomal complex with human ZNF277 that competes with the PRMT3–uS5 interaction

Dionne

Bergeron

Landry-Voyer

et al. 2019

Journal of Biological Chemistry

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Edited by Karin Musier-ForsythRibosomal (r)-proteins are generally viewed as ubiquitous, constitutive proteins that simply function to maintain ribosome integrity. However, findings in the past decade have led to the idea that r-proteins have evolved specialized functions beyond the ribosome. For example, the 40S ribosomal protein uS5 (RPS2) is known to form an extraribosomal complex with the protein arginine methyltransferase PRMT3 that is conserved from fission yeast to humans. However, the full scope of uS5's extraribosomal functions, including whether uS5 interacts with any other proteins, is not known. In this study, we identify the conserved zinc finger protein 277 (ZNF277) as a new uS5-associated protein by using quantitative proteomics approaches in human cells. As previously shown for PRMT3, we found that ZNF277 uses a C2H2-type zinc finger domain to recognize uS5. Analysis of protein-protein interactions in living cells indicated that the ZNF277-uS5 complex is found in the cytoplasm and the nucleolus. Furthermore, we show that ZNF277 and PRMT3 compete for uS5 binding, because overexpression of PRMT3 inhibited the formation of the ZNF277-uS5 complex, whereas depletion of cellular ZNF277 resulted in increased levels of uS5-PRMT3. Notably, our results reveal that ZNF277 recognizes nascent uS5 in the course of mRNA translation, suggesting cotranslational assembly of the ZNF277-uS5 complex. Our findings thus unveil an intricate network of evolutionarily conserved protein-protein interactions involving extraribosomal uS5, suggesting a key role for uS5 beyond the ribosome. . 2 The abbreviations used are: r-protein, ribosomal protein; ZNF277, zinc finger protein 277; PRMT3, protein arginine methyltransferase 3; uS5/RPS2, 40S ribosomal protein S2; ZF, zinc finger; SILAC, stable isotope labeling by amino acids in cell culture; PDCD2, programmed cell death protein 2; PDCD2L, PDCD2-like protein; AP, affinity purification; BiFC, bimolecular fluorescence complementation; YFP, yellow fluorescent protein; qPCR, quantitative PCR; MCS, multiple cloning site.

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Section: C2h2 Zinc Finger Domains Of Znf277 Are Important For the Assmentioning

confidence: 80%

The 40S ribosomal protein uS5 (RPS2) assembles into an extraribosomal complex with human ZNF277 that competes with the PRMT3–uS5 interaction

Dionne

Bergeron

Landry-Voyer

et al. 2019

Journal of Biological Chemistry

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“…2A, lanes 1, 3, 5, and 6). Kap123 is considered the primary importin for RPs (22), while Kap104 has been shown to interact with specific RPs (16,18,20,23). This result suggests that the Rps2 core is needed for the recruitment of an importin.…”

mentioning

confidence: 99%

Tsr4 Is a Cytoplasmic Chaperone for the Ribosomal Protein Rps2 in Saccharomyces cerevisiae

Black

Musalgaonkar

Johnson

2019

Molecular and Cellular Biology

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Eukaryotic ribosome biogenesis requires the action of approximately 200 trans-acting factors and the incorporation of 79 ribosomal proteins (RPs). The delivery of RPs to preribosomes is a major challenge for the cell because RPs are often highly basic and contain intrinsically disordered regions prone to nonspecific interactions and aggregation. To counteract this, eukaryotes developed dedicated chaperones for certain RPs that promote their solubility and expression, often by binding eukaryote-specific extensions of the RPs. Rps2 (uS5) is a universally conserved RP that assembles into nuclear pre-40S subunits. However, a chaperone for Rps2 had not been identified. Our laboratory previously characterized Tsr4 as a 40S biogenesis factor of unknown function. Here, we report that Tsr4 cotranslationally associates with Rps2. Rps2 harbors a eukaryote-specific N-terminal extension that is critical for its interaction with Tsr4. Moreover, Tsr4 perturbation resulted in decreased Rps2 levels and phenocopied Rps2 depletion. Despite Rps2 joining nuclear pre-40S particles, Tsr4 appears to be restricted to the cytoplasm. Thus, we conclude that Tsr4 is a cytoplasmic chaperone dedicated to Rps2.

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“…We found that binding of importin α (Kap60 in yeast) competed with Yar1 for binding to Rps3, suggesting that one N-domain of Rps3 can only bind either importin or Yar1 (Figure 1, Step III). As we nevertheless detected complexes containing Rps3 and both Yar1 and Kap60 in vivo , Rps3-dimers with Yar1 bound to one and importin bound to the second Rps3 N-domain might represent the preferred conformation for nuclear import 5. It is still a puzzle how such architecture is retained, but assuming that the complex is imported immediately after one of the two Yar1 copies has been replaced by importin, the limited time for a second importin to access the second Rps3 copy might make it simply more likely that one Yar1 is maintained.…”

mentioning

confidence: 67%

“…NLSs comprise the binding sites for importins, which mediate the transport of proteins through the nuclear pores into the nucleus 6. Our analyses suggest that several redundant routes engage in Rps3 import, with major contribution from the classical importin α/importin β pathway 5. We found that binding of importin α (Kap60 in yeast) competed with Yar1 for binding to Rps3, suggesting that one N-domain of Rps3 can only bind either importin or Yar1 (Figure 1, Step III).…”

mentioning

confidence: 87%

When a ribosomal protein grows up - the ribosome assembly path of Rps3

Pertschy¹

2017

Microb Cell

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The biogenesis of ribosomes is a central process in all dividing cells. Eukaryotic ribosomes are composed of a large 60S and a small 40S subunit, each comprising a complex assembly of ribosomal RNA (rRNA) and ribosomal proteins (r-proteins). The synthesis of these constituents is spatially separated, with r-proteins being produced by translation in the cytoplasm, while rRNA is generated by transcription in the nucleus. Hence, the arrangement of r-proteins and rRNA into large ribonucleoprotein complexes requires dedicated mechanisms ensuring their encounter in the same compartment. To this end, r-proteins need to be safely delivered to the nucleus where they assemble with the rRNA. Beyond these initial challenges, the synthesis of ribosomes does not merely comprise the joining of r-proteins with rRNA, but occurs in a complex assembly line involving multiple maturation steps, including the processing and folding of rRNA. R-proteins usually have composite rRNA binding sites, with several different rRNA helices contributing to the full interaction. Not all of these interaction sites may already be accessible at the point when an r-protein is incorporated, necessitating that some of the r-protein-rRNA contacts are formed at later maturation stages. In our two recent studies, we investigated the ribosome assembly path of r-proteins in the yeast Saccharomyces cerevisiae using the small subunit r-protein S3 (Rps3) as a model. Our studies revealed intricate mechanisms to protect the protein, transport it into the nucleus, integrate it into pre-ribosomal precursor particles and promote its final stable association with 40S subunits.

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Nuclear import of dimerized ribosomal protein Rps3 in complex with its chaperone Yar1

Cited by 28 publications

References 46 publications

The 40S ribosomal protein uS5 (RPS2) assembles into an extraribosomal complex with human ZNF277 that competes with the PRMT3–uS5 interaction

The 40S ribosomal protein uS5 (RPS2) assembles into an extraribosomal complex with human ZNF277 that competes with the PRMT3–uS5 interaction

Tsr4 Is a Cytoplasmic Chaperone for the Ribosomal Protein Rps2 in Saccharomyces cerevisiae

When a ribosomal protein grows up - the ribosome assembly path of Rps3

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