| 2017 |
Yeast Rps26 contributes to mRNA-specific translation by recognition of the Kozak sequence in well-translated mRNAs; Rps26-deficient ribosomes preferentially translate mRNAs from select stress-response pathways. Exposure to stress leads to formation of Rps26-deficient ribosomes and increased translation of their target mRNAs, establishing a feed-forward translational stress response loop. |
Separation of two ribosome populations (with and without Rps26) from same cells combined with RNA-seq; genetic and biochemical analysis in S. cerevisiae |
Nature structural & molecular biology |
High |
28759050
|
| 2022 |
The chaperone Tsr2 releases Rps26 from fully assembled ribosomes in the presence of high Na+ or elevated pH in vitro, and is required for Rps26 release in vivo during osmotic/pH stress. Tsr2 stores free Rps26 and promotes its reincorporation into ribosomes after stress subsides, enabling a reversible ribosome population change. A DBA-associated residue in Rps26 mediates the Na+ effect. |
In vitro biochemical assay (Tsr2-mediated Rps26 release), in vivo yeast genetics (Tsr2 deletion), ribosome fractionation, site-directed mutagenesis |
Science advances |
High |
35213229
|
| 2024 |
Released Rps26 (from the Rps26•Tsr2 complex during high-salt stress) is degraded via the Pro/N-degron pathway. The GID-complex E3 ubiquitin ligase and its adaptor Gid4 mediate polyubiquitination of Rps26 at Lys66 and Lys70, enabling Tsr2 recycling, accumulation of Rps26-deficient ribosomes to ~50% of total, and high-salt stress resistance. |
Yeast genetics (N-terminal proline substitution, GID-complex/Gid4 deletion), ubiquitination assays, polysome profiling, salt-stress phenotype assays |
bioRxivpreprint |
High |
|
| 2013 |
RPS26 knockdown induces p53 stabilization via an RPL11-dependent mechanism. RPS26 protein interacts with Mdm2 and inhibits Mdm2-mediated p53 ubiquitination. RPS26 also interacts with p53 independently of Mdm2 and co-exists in a complex with p53 and p300. RPS26 knockdown impairs p53 transcriptional activity, p53 acetylation, and recruitment of p53 to target gene promoters in response to DNA damage, abolishing G2/M arrest. |
siRNA knockdown in cells, Co-immunoprecipitation, p53 ubiquitination assay, ChIP assay, cell cycle analysis |
Oncogene |
Medium |
23728348
|
| 2013 |
Upf1 (NMD regulator) interacts specifically with Rps26 of the 40S ribosomal subunit. This interaction is mediated by the N-terminal CH domain of Upf1, is dependent on ATP, and occurs without simultaneous association of eRF1, eRF3, Upf2, or Upf3. |
Two-hybrid screen, in vitro binding assay, coimmunoprecipitation with epitope-tagged 40S subunits, domain mapping with UPF1 mutations |
RNA (New York, N.Y.) |
Medium |
23801788
|
| 2010 |
Mutations in RPS26 cause Diamond-Blackfan anemia (DBA). Lymphoblastoid cells from patients with RPS26 mutations show elevated 18S-E pre-rRNA, and siRNA knockdown of RPS26 in HeLa cells phenocopies this, indicating RPS26 is required for 18S rRNA processing at the 18S-E step. |
Pre-rRNA analysis in patient lymphoblastoid cells; siRNA knockdown in HeLa cells with pre-rRNA northern blot |
American journal of human genetics |
High |
20116044
|
| 2005 |
Human RPS26 protein binds to the first intron and mRNA fragment of its own pre-mRNA (detected by nitrocellulose filtration). Recombinant RPS26 suppresses in vitro splicing of both conventional and alternative RPS26 mRNAs. Toe-printing mapped RPS26 binding to two clusters in the pre-mRNA secondary structure flanking the conventional and alternative 3' splice sites, establishing an autoregulatory feedback mechanism at the level of pre-mRNA splicing. |
Nitrocellulose filtration binding assay, in vitro splicing assay with recombinant protein and HeLa nuclear extract, toe-printing |
Biochimica et biophysica acta |
Medium |
15716004
|
| 2011 |
The eukaryote-specific motif YxxPKxYxK (fragment 60–71) of human rpS26e directly contacts mRNA at positions -3 to -9 relative to the E-site codon on the 80S ribosome, establishing this region as part of the mRNA binding channel 5' of the E site. X-ray structural analysis showed this motif is not involved in intraribosomal contacts, implying its function in translation. |
Site-directed cross-linking with mRNA analogues bearing perfluorophenyl azide at defined positions, proteolytic mapping of cross-linked peptides in human 80S ribosomal complexes, X-ray structural analysis of Tetrahymena thermophila 40S subunit |
Nucleic acids research |
High |
22167470
|
| 2023 |
RPS26 C-terminal domain binds mRNA at positions -10 to -16 (AUG upstream nucleotides at the exit channel). This binding exerts positive effects on Kozak-driven translation and negative effects on TISU-driven translation. CRISPR-Cas9 mutation of the RPS26 C-terminus (RPS26dC) confers resistance to glucose starvation and mTOR inhibition, reduces basal mTOR activity, and activates AMPK, linking RPS26 C-terminal RNA binding to energy metabolism and translational stress responses. |
CRISPR-Cas9 mutagenesis of RPS26 C-terminus, translatome analysis (ribosome profiling/RNA-seq), reporter assays with Kozak/TISU elements, glucose starvation and mTOR inhibition assays, AMPK/mTOR activity measurements |
Nucleic acids research |
High |
37013984
|
| 2016 |
The eukaryote-specific Y62-K70 (YxxPKxYxK) segment of Rps26 is essential for viability (complete deletion is lethal), and simultaneous alanine substitution of five conserved residues within this segment causes growth defects and accumulation of free 60S subunits (indicating impaired 80S assembly). Single-amino-acid substitutions in this motif did not affect function. Human Rps26 expressed in yeast supports growth but causes altered 40S/60S ratios, indicating a role for Rps26 in 40S subunit assembly and 80S ribosome formation rather than specifically in translation initiation. |
Alanine-scanning mutagenesis in S. cerevisiae, polysome profiling, growth assays under stress conditions, complementation with human RPS26 |
mSphere |
Medium |
27303706
|
| 2022 |
The eukaryote-specific YxxPKxYxK motif of human eS26 contacts the same mRNA nucleotide residues as translation initiation factor eIF3 on mammalian 80S ribosomes. Simultaneous replacement of all five conserved residues (5A mutant) increases the light polysome fraction and enhances eIF3e content in that fraction, suggesting involvement of this motif in fine-tuning selective translation. |
Site-directed cross-linking on mammalian 80S ribosomes, transfection of HEK293T cells with FLAG-tagged wild-type or mutant eS26 constructs, polysome profiling with western blot analysis, real-time PCR |
Biochimica et biophysica acta. Gene regulatory mechanisms |
Medium |
35817369
|
| 2018 |
The C-terminus of eS26 is required for nucleophosmin binding to the 40S ribosomal subunit. FLAG-tagged eS26 incorporates into 40S subunits without affecting ribosome assembly or translational activity, but eS26FLAG-containing ribosome fractions show reduced nucleophosmin content. Direct binding of nucleophosmin to isolated eS26 and to 40S subunits was demonstrated with recombinant protein in the presence of HeLa nuclear extract (which phosphorylates nucleophosmin), implicating the eS26 C-terminus in the mRNA exit site region for nucleophosmin-dependent nuclear export of pre-40S subunits. |
Doxycycline-inducible expression of C-terminally FLAG-tagged eS26 in HEK293-derived cells, polysome fractionation with western blot, in vitro binding assay with recombinant nucleophosmin and HeLa nuclear extract |
Biochimica et biophysica acta. Proteins and proteomics |
Medium |
29563070
|
| 1984 |
Ribosomal protein S26 is located at the mRNA binding site of rat liver ribosomes, as established by affinity labeling with a radioactive alkylating heptauridylate derivative; labeling was abolished by competing poly(U). |
Affinity labeling with radioactive alkylating oligonucleotide analogue on rat liver ribosomes; competition with poly(U) |
Molecular biology reports |
Medium |
6708946
|
| 2004 |
mRNA positions -4 to -9 (5' of the E-site codon) on the human 80S ribosome are primarily cross-linked to protein S26, establishing S26 as the principal ribosomal protein neighboring mRNA 5' of the E site codon. |
Site-directed UV cross-linking with photoactivatable mRNA analogues positioned in human 80S ribosomal complexes; protein identification by gel analysis |
Molekuliarnaia biologiia |
Medium |
15612591
|
| 2006 |
RPS26A (but not RPS26B) in S. cerevisiae is required for FLO11-mediated haploid adhesive and diploid pseudohyphal growth. FLO11-lacZ activity is absent in rps26AΔ strains despite normal FLO11 mRNA levels, indicating that Rps26 amount is critical for accurate translation of FLO11 mRNA and the dimorphic switch. Overexpression of RPS26B or RPS26B driven by the RPS26A promoter complements this defect, indicating the proteins are functionally interchangeable when expressed at sufficient levels. |
Yeast genetics (single and double deletion strains), reporter assays (FLO11-lacZ), northern blot for FLO11 mRNA, complementation experiments |
Molecular genetics and genomics |
Medium |
16721598
|
| 2025 |
Depletion of RPS26 (and its chaperone TSR2) modulates FMRpolyG production from CGG-repeat-expanded FMR1 mRNA (repeat-associated non-AUG translation). RPS26 was identified as enriched on CGG-expanded FMR1 RNA by RNA-tagging/mass spectrometry. RPS26 insufficiency preferentially impacts translation of mRNAs with short, GC-rich 5'UTRs. |
RNA-tagging and mass spectrometry screening for proteins enriched on CGGexp FMR1 RNA; siRNA knockdown of RPS26 and TSR2; FMRpolyG production assay; translatome analysis |
eLife |
Medium |
40377206
|
| 2025 |
Tissue-specific patterns of ribosome termination pausing correlate with the stoichiometry of Rps26, which modulates mRNA:rRNA interactions at the stop codon. Reduced Rps26 levels are associated with altered termination pausing and increased stop codon slippage. |
Terminating ribosome profiling in mammalian cells; massively parallel reporter assays; correlation of Rps26 stoichiometry with termination pausing patterns across tissues |
bioRxivpreprint |
Low |
|
| 2024 |
Cryo-EM structures of archaeal (Saccharolobus solfataricus) small ribosomal subunit initiation complexes show archaeal eS26 positioned in the mRNA exit channel wrapped around the 3' end of ribosomal RNA, as in eukaryotes, and its position is incompatible with an SD:antiSD duplex in the exit channel, suggesting a conserved role of eS26 in translation regulation. |
Cryo-EM structure determination of archaeal ribosomal initiation complexes |
bioRxivpreprint |
Medium |
|
| 2022 |
RPS26 deficiency in human erythroid progenitor cells (HUDEP-1) causes imbalanced ribosomal RNA production, upregulation of pro-apoptotic genes, reduced cell viability, increased intracellular calcium, and impaired erythroid differentiation. |
siRNA knockdown of RPS26 in HUDEP-1 cells; rRNA analysis; flow cytometry for erythroid markers and apoptosis; calcium measurement |
Frontiers in genetics |
Medium |
36579335
|
| 2018 |
Specific knockout of Rps26 in mouse oocytes arrests chromatin configuration at the NSN-to-SN transition, decreases mRNA transcription, reduces H3K4/H3K9 trimethylation and DNA methylation, lowers oocyte-derived growth factors (GDF9, BMP15, CX37), and disrupts PI3K/AKT/FOXO3a signaling, causing premature ovarian failure. |
Conditional knockout mouse (oocyte-specific Rps26 deletion), histology, immunofluorescence for histone modifications and DNA methylation, western blot for signaling pathway components |
Cell death & disease |
Medium |
30451825
|