Affinage

RPL35

Large ribosomal subunit protein uL29 · UniProt P42766

Length
123 aa
Mass
14.6 kDa
Annotated
2026-06-10
50 papers in source corpus 14 papers cited in narrative 14 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RPL35 (uL29/L35) is a structural protein of the 60S large ribosomal subunit that is also required for early large-subunit biogenesis (PMID:863909, PMID:20392820). First purified from rat liver ribosomes and sequenced as a 122-residue protein related to archaebacterial L33/L29E and E. coli L29 (PMID:863909, PMID:2322279), it assembles in the nucleolus on early pre-60S particles, and its depletion in yeast causes a deficit of 60S subunits, a strong delay in 27SB-to-7S pre-rRNA processing at ITS2 site C2, and a block in pre-60S nucleolus-to-nucleoplasm release (PMID:20392820). In eukaryotes the protein carries a 54-aa C-terminal expansion segment whose first 25 residues are essential for nuclear import while its last 29 residues mediate ribosome docking to the endoplasmic reticulum (PMID:18523488). RPL35 directly binds elongation factor eEF2 to control translational elongation and ER-localized secretion of nascent protein (PMID:26297660). Beyond the ribosome, RPL35 has acquired extra-ribosomal regulatory roles: it binds the lncRNA lncNB1 to enhance E2F1 translation and drive a DEPDC1B–ERK–N-Myc axis supporting neuroblastoma proliferation (PMID:31690716), it acts as an antiviral host factor that recruits the E3 ligase AMFR to mediate K48-linked polyubiquitination and proteasomal degradation of the FMDV capsid protein VP2 (PMID:41065387), and its own stability is set by competing factors—negatively by TENT5A within an EGR1/TENT5A/RPL35/mTOR axis (PMID:39570560) and positively by GADD45GIP1, which blocks ubiquitin-mediated degradation of RPL35 and thereby restrains the PERK–eIF2α ER stress pathway (PMID:40604925). RPL35 also protects chondrocytes against senescence and cartilage degeneration via suppression of the Hedgehog/Gli1 pathway (PMID:38596341).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1977 High

    Established that L35 is a discrete, purifiable protein component of the eukaryotic 60S large ribosomal subunit, defining the molecule biochemically.

    Evidence Ion-exchange/gel-filtration purification, SDS-PAGE and amino acid composition from rat liver ribosomes

    PMID:863909

    Open questions at the time
    • No sequence or structural position within the subunit
    • No functional role assigned
  2. 1990 High

    Resolved the primary structure and evolutionary relationships of the protein, placing it in the L29/L33E family and revealing an internal sequence duplication.

    Evidence cDNA and N-terminal protein sequencing with Southern blotting

    PMID:2322279

    Open questions at the time
    • Functional significance of the internal duplication unknown
    • No mechanistic role established
  3. 2008 Medium

    Showed that the eukaryote-specific C-terminal expansion segment partitions distinct trafficking functions—nuclear import versus ER docking—separable from core ribosomal/translational function.

    Evidence Recombinant ribosome analysis with deletion mutagenesis and subcellular localization assays

    PMID:18523488

    Open questions at the time
    • Import receptor not identified here
    • Single lab; structural basis of ER docking not resolved
  4. 2010 High

    Defined the precise biogenesis role: L35 assembles in the nucleolus on early pre-60S particles and is required for 27SB pre-rRNA processing and pre-60S nuclear export.

    Evidence Yeast depletion with pulse-chase, northern, primer extension and flow cytometry

    PMID:20392820

    Open questions at the time
    • Direct interaction partners on pre-60S not mapped
    • Human ortholog biogenesis role inferred from yeast
  5. 2015 Medium

    Connected RPL35 to the elongation machinery and secretory output by demonstrating a direct eEF2 interaction controlling elongation and ER targeting of a secreted protein.

    Evidence Reciprocal Co-IP, FRET, colocalization and mutagenesis/knockdown in bovine mammary epithelial cells

    PMID:26297660

    Open questions at the time
    • Generality beyond CSN2 substrate not tested
    • Mechanism of methionine-dependent regulation unclear
  6. 2019 Medium

    Revealed an extra-ribosomal, RNA-binding function in which RPL35 partners with lncNB1 to selectively enhance E2F1 translation and drive an oncogenic ERK/N-Myc cascade.

    Evidence RNA pulldown, Co-IP, knockdown, reporter assays and in vivo tumor regression in neuroblastoma

    PMID:31690716

    Open questions at the time
    • Whether translational selectivity is ribosome-dependent unclear
    • Direct lncNB1-binding region of RPL35 not mapped
  7. 2021 Low

    Extended the neuroblastoma role to metabolic control, placing RPL35 upstream of an ERK/HIF1α axis governing aerobic glycolysis.

    Evidence siRNA knockdown with metabolic assays, Western blot and qPCR

    PMID:34873488

    Open questions at the time
    • Pathway placement inferred without epistasis rescue
    • Single lab, correlative
  8. 2021 Low

    Implicated RPL35 as a host target of a bacterial adhesin, linking the protein to proliferative signaling during Mycoplasma infection.

    Evidence Phage display, far-Western, colocalization, TMT proteomics and MTT in urothelial cells

    PMID:34832605

    Open questions at the time
    • Downstream effects correlative
    • No reciprocal validation of functional consequence
  9. 2021 Medium

    Provided structural ligand-binding information by mapping small-molecule (atazanavir, artesunate) contact sites on human RPL35.

    Evidence Molecular docking and NMR chemical-shift titration

    PMID:33823521

    Open questions at the time
    • Functional consequence of binding not tested in vivo
    • Single study
  10. 2022 Low

    Identified DDX10 as a physical interactor of RPL35 in colorectal cancer, hinting at a link between RPL35 and splicing-associated processes.

    Evidence LC-MS/MS and Co-IP

    PMID:35109823

    Open questions at the time
    • No mechanistic follow-up for RPL35 specifically
    • Single Co-IP without reciprocal validation
  11. 2024 Medium

    Established a protective role for RPL35 in chondrocyte homeostasis, where its loss promotes senescence via Hedgehog/Gli1 activation and its restoration limits OA cartilage degeneration.

    Evidence RNA-seq, siRNA knockdown, adenoviral overexpression and an in vivo mouse OA model

    PMID:38596341

    Open questions at the time
    • Mechanism linking RPL35 to Hh/Gli1 not defined
    • Single lab
  12. 2024 Medium

    Defined RPL35 as a regulated node whose stability is negatively controlled by TENT5A within an EGR1/TENT5A/RPL35 axis that restrains mTOR signaling in hepatocellular carcinoma.

    Evidence Co-IP, GST pull-down, mass spectrometry, ChIP and luciferase reporter with gain/loss-of-function

    PMID:39570560

    Open questions at the time
    • Mechanism of TENT5A-mediated destabilization unclear
    • Direct link of RPL35 to mTOR not resolved
  13. 2025 Medium

    Showed RPL35 protein abundance is set by ubiquitin-mediated turnover that GADD45GIP1 antagonizes, coupling RPL35 stability to control of the PERK-eIF2α ER stress response.

    Evidence IP/LC-MS/MS, siRNA knockdown, overexpression rescue and ubiquitination/PERK/eIF2α Western blots in osteosarcoma

    PMID:40604925

    Open questions at the time
    • E3 ligase for RPL35 in this context not identified
    • Single lab
  14. 2025 Medium

    Demonstrated an antiviral effector function: RPL35 recruits AMFR to ubiquitinate and degrade the FMDV capsid protein VP2, a defense the virus evades by blocking RPL35 nuclear translocation.

    Evidence Co-IP, proteomic screen, ubiquitination assay, VP2-K217R recombinant virus and KPNA3 degradation assay

    PMID:41065387

    Open questions at the time
    • Whether antiviral activity is ribosome-independent unclear
    • Not independently replicated

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RPL35's core ribosomal function is mechanistically partitioned from its diverse extra-ribosomal roles—and what governs its differential engagement across these contexts—remains unresolved.
  • No structural model integrating ribosomal and extra-ribosomal interfaces
  • Direct RNA- and protein-binding surfaces for extra-ribosomal partners unmapped
  • No human Mendelian disease link established in the corpus

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005198 structural molecule activity 3 GO:0003723 RNA binding 1 GO:0045182 translation regulator activity 1
Localization
GO:0005634 nucleus 2 GO:0005783 endoplasmic reticulum 2 GO:0005840 ribosome 2 GO:0005730 nucleolus 1
Pathway
R-HSA-1852241 Organelle biogenesis and maintenance 1 R-HSA-392499 Metabolism of proteins 1 R-HSA-8953854 Metabolism of RNA 1
Complex memberships
60S large ribosomal subunitpre-60S ribosomal particle

Evidence

Reading pass · 14 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1977 RPL35 (L35) was isolated and purified as a component of the 60S large ribosomal subunit from rat liver ribosomes, with molecular weight estimated by SDS-PAGE and amino acid composition determined. Ion exchange chromatography, gel filtration, SDS-PAGE, amino acid composition analysis The Journal of biological chemistry High 863909
1990 The primary structure of rat ribosomal protein L35 was determined: 122 amino acids (N-terminal methionine removed post-translationally), molecular weight 14,412 Da; the protein is related to archaebacterial L33/L29E and E. coli L29, and contains a possible internal duplication of 11 residues. cDNA sequencing, N-terminal amino acid sequencing, Southern blotting Biochemical and biophysical research communications High 2322279
2008 Human RPL35 carries a 54 aa eukaryotic expansion segment (ES) at the C-terminal end. The first 25 aa of the ES are essential for nuclear import of the protein. The last 29 residues of the ES are not required for ribosome structural or translational functions but serve as a contact site for ribosomal docking to the endoplasmic reticulum. In vivo recombinant ribosome analysis, deletion mutagenesis, subcellular localization assays Biochemistry and cell biology Medium 18523488
2010 In yeast (Saccharomyces cerevisiae), depletion of ribosomal protein L35 results in a deficit in 60S ribosomal subunits, half-mer polysomes, and a strong delay in processing of 27SB to 7S pre-rRNAs (cleavage at ITS2 site C2). L35 assembles in the nucleolus and associates with early pre-60S ribosomal particles. Loss of L35 also blocks release of pre-60S particles from nucleolus to nucleoplasm and mildly delays G1 phase of the cell cycle. In vivo depletion, pulse-chase labeling, northern hybridization, primer extension, flow cytometry, deletion mutant analysis Nucleic acids research High 20392820
2015 RPL35 controls ribosomal translational elongation of β-casein (CSN2) by directly interacting with eukaryotic translational elongation factor 2 (eEF2), with eEF2 acting downstream of RPL35. RPL35 also controls secretion of CSN2 by localizing it to the ER. Methionine promotes RPL35 expression, and RPL35 mediates the methionine signal to regulate CSN2 expression in bovine mammary epithelial cells. Co-immunoprecipitation, colocalization, FRET, gene mutation/overexpression/knockdown Archives of biochemistry and biophysics Medium 26297660
2019 RPL35 binds the long noncoding RNA lncNB1, and this interaction enhances E2F1 protein synthesis (translational), leading to DEPDC1B gene transcription. DEPDC1B (GTPase-activating protein) then induces ERK phosphorylation and N-Myc protein stabilization. RPL35 knockdown reduces neuroblastoma cell clonogenic capacity. RNA pulldown, Co-IP, loss-of-function (knockdown), reporter assays, in vivo tumor regression Nature communications Medium 31690716
2021 RPL35 knockdown in neuroblastoma cells decreases aerobic glycolysis and increases mitochondrial respiration, decreases HIF1α expression (mRNA and protein), and negatively regulates ERK pathway activity. RPL35 modulates aerobic glycolysis in part through the RPL35/ERK/HIF1α axis. siRNA knockdown, Western blot, metabolic assays (glycolysis/respiration), qPCR American journal of cancer research Low 34873488
2021 RPL35 interacts directly with the Mycoplasma genitalium adhesin protein MgPa (verified by far-Western blot and co-localization), and this interaction promotes expression of EIF2, SRP68, SERBP1, RPL35A, EGF, and TGF-β, and enhances urothelial cell proliferation. T7 phage-display library screening, far-Western blot, co-localization, TMT proteomics, qRT-PCR, MTT assay Pathogens Low 34832605
2021 Drug development study using NMR titration identified that atazanavir and artesunate bind to human RPL35 near the N-terminus; NMR chemical shift mapping revealed the amino acids at the ligand-binding sites, including a binding cluster accessible when RPL35 is integrated into the ribosome. Molecular docking, NMR spectroscopy (chemical shift titration) Skin pharmacology and physiology Medium 33823521
2022 DDX10 interacts with RPL35 (identified by LC-MS/MS and confirmed by Co-IP), and DDX10 is proposed to regulate CRC cell proliferation and metastasis via splicing-related mechanisms involving RPL35. LC-MS/MS, Co-IP Cancer cell international Low 35109823
2024 RPL35 protein expression is reduced in chondrocytes under excessive mechanical loading. RPL35 knockdown alone induces chondrocyte senescence, decreases anabolic markers, and increases catabolic markers in part through activation of the Hedgehog (Hh)/Gli1 pathway. Intra-articular overexpression of RPL35 protects against cartilage degeneration in a mouse OA model. RNA sequencing, siRNA knockdown, adenovirus-mediated overexpression, Western blot, immunostaining, in vivo mouse OA model Journal of orthopaedic translation Medium 38596341
2024 TENT5A interacts with RPL35 (Co-IP + GST pull-down confirmed by mass spectrometry) and negatively regulates RPL35 protein stability. The EGR1/TENT5A/RPL35 axis is involved in ribosome biogenesis and negatively regulates the mTOR pathway in hepatocellular carcinoma cells. Co-immunoprecipitation, GST pull-down, mass spectrometry, chromatin immunoprecipitation, dual-luciferase reporter assay, gain/loss-of-function experiments Cellular oncology Medium 39570560
2025 GADD45GIP1 interacts with RPL35 (identified by IP/LC-MS/MS, ranked second among 263 interacting proteins) and inhibits ubiquitin-mediated proteasomal degradation of RPL35. RPL35 knockdown activates the PERK-eIF2α ER stress pathway, increases ER stress, and reduces osteosarcoma cell viability. Overexpression of RPL35 counteracts the decrease in cell viability caused by GADD45GIP1 knockdown. Immunoprecipitation, LC-MS/MS, siRNA knockdown, overexpression rescue, Western blot for ubiquitination/PERK/eIF2α Cancer cell international Medium 40604925
2025 RPL35 functions as an antiviral host protein against FMDV by directly interacting with the FMDV structural protein VP2 and recruiting the E3 ubiquitin ligase AMFR to mediate K48-linked polyubiquitination and proteasomal degradation of VP2. The Lys217 residue of VP2 is critical for this RPL35-mediated antiviral activity. FMDV counteracts this by inducing degradation of KPNA3, thereby blocking RPL35 nuclear translocation. Co-IP, proteomic screen, ubiquitination assay, site-directed mutagenesis (VP2-K217R recombinant virus), KPNA3 knockdown/degradation assay Journal of virology Medium 41065387

Source papers

Stage 0 corpus · 50 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2000 Construction, phenotypic analysis, and immunogenicity of a UL5/UL29 double deletion mutant of herpes simplex virus 2. Journal of virology 96 10933704
2019 The long noncoding RNA lncNB1 promotes tumorigenesis by interacting with ribosomal protein RPL35. Nature communications 84 31690716
1976 Isolation of eukaryotic ribosomal proteins. Purification and characterization of the 60 S ribosomal subunit proteins L4, L5, L7, L9, L11, L12, L13, L21, L22, L23, L26, L27, L30, L33, L35', L37, and L39. The Journal of biological chemistry 70 1002715
1977 Isolation of eukaryotic ribosomal proteins. Purification and characterization of 60 S ribosomal subunit proteins L3, L6, L7', L8, L10, L15, L17, L18, L19, L23', L25, L27', L28, L29, L31, L32, L34, L35, L36, L36', and L37'. The Journal of biological chemistry 68 863909
2010 Human cytomegalovirus UL29/28 protein interacts with components of the NuRD complex which promote accumulation of immediate-early RNA. PLoS pathogens 64 20585571
1992 Messenger RNA secondary structure and translational coupling in the Escherichia coli operon encoding translation initiation factor IF3 and the ribosomal proteins, L35 and L20. Journal of molecular biology 58 1453449
2010 Ribosomal protein L35 is required for 27SB pre-rRNA processing in Saccharomyces cerevisiae. Nucleic acids research 47 20392820
2005 R-state haemoglobin with low oxygen affinity: crystal structures of deoxy human and carbonmonoxy horse haemoglobin bound to the effector molecule L35. Journal of molecular biology 40 16403522
2009 Human cytomegalovirus UL28 and UL29 open reading frames encode a spliced mRNA and stimulate accumulation of immediate-early RNAs. Journal of virology 33 19625400
2002 Translational feedback regulation of the gene for L35 in Escherichia coli requires binding of ribosomal protein L20 to two sites in its leader mRNA: a possible case of ribosomal RNA-messenger RNA molecular mimicry. RNA (New York, N.Y.) 31 12166643
1996 Suppression of a sensor kinase-dependent phenotype in Pseudomonas syringae by ribosomal proteins L35 and L20. Journal of bacteriology 30 8626280
2003 Mutation of single hydrophobic residue I27, L35, F39, L58, L65, L67, or L71 in the N terminus of VP5 abolishes interaction with the scaffold protein and prevents closure of herpes simplex virus type 1 capsid shells. Journal of virology 25 12634364
2022 DDX10 promotes the proliferation and metastasis of colorectal cancer cells via splicing RPL35. Cancer cell international 24 35109823
2015 60S ribosomal protein L35 regulates β-casein translational elongation and secretion in bovine mammary epithelial cells. Archives of biochemistry and biophysics 24 26297660
1996 A long-range RNA-RNA interaction forms a pseudoknot required for translational control of the IF3-L35-L20 ribosomal protein operon in Escherichia coli. The EMBO journal 23 8861967
2010 Porcine circovirus type 2 morphogenesis in a clone derived from the l35 lymphoblastoid cell line. Journal of comparative pathology 22 20800239
1994 Phorbol ester induction of differentiation and apoptosis in the K562 cell line is accompanied by marked decreases in the stability of globin mRNAs and decreases in the steady state level of mRNAs encoding for ribosomal proteins L35, L31, L27, and L21. Cellular & molecular biology research 20 7804323
2022 18β-glycyrrhetinic acid regulates mitochondrial ribosomal protein L35-associated apoptosis signaling pathways to inhibit proliferation of gastric carcinoma cells. World journal of gastroenterology 19 35979263
2011 RNase Y is responsible for uncoupling the expression of translation factor IF3 from that of the ribosomal proteins L35 and L20 in Bacillus subtilis. Molecular microbiology 18 21843271
2007 A competition mechanism regulates the translation of the Escherichia coli operon encoding ribosomal proteins L35 and L20. Journal of molecular biology 17 18037435
2008 The role of expansion segment of human ribosomal protein L35 in nuclear entry, translation activity, and endoplasmic reticulum docking. Biochemistry and cell biology = Biochimie et biologie cellulaire 15 18523488
2021 RPL35 promotes neuroblastoma progression via the enhanced aerobic glycolysis. American journal of cancer research 14 34873488
2020 Herpes Simplex Virus Type 1 Clinical Isolates Respond to UL29-Targeted siRNA Swarm Treatment Independent of Their Acyclovir Sensitivity. Viruses 13 33322225
1996 Rat hepatoma L35 cells, a liver-differentiated cell line, display resistance to bile acid repression of cholesterol 7 alpha-hydroxylase. Journal of lipid research 12 8728321
1997 Transcriptional induction of cholesterol 7alpha-hydroxylase by dexamethasone in L35 hepatoma cells requires sulfhydryl reducing agents. The Journal of biological chemistry 11 9006961
1990 Ribosomal protein L35: identification in spinach chloroplasts and isolation of a cDNA clone encoding its cytoplasmic precursor. Biochemistry 11 2271612
2024 RPL35 downregulated by mechanical overloading promotes chondrocyte senescence and osteoarthritis development via Hedgehog-Gli1 signaling. Journal of orthopaedic translation 10 38596341
1997 Sequence analysis of the bovine herpesvirus type 1 genes homologous to the DNA polymerase (UL30), the major DNA-binding protein (UL29) and ICP18.5 assembly protein (UL28) genes of herpes simplex virus. Archives of virology 10 9155875
1990 The primary structure of rat ribosomal protein L35. Biochemical and biophysical research communications 10 2322279
2023 Single therapeutic dose of an antiviral UL29 siRNA swarm diminishes symptoms and viral load of mice infected intranasally with HSV-1. Smart medicine 9 39188276
2021 Drug Development for Target Ribosomal Protein rpL35/uL29 for Repair of LAMB3R635X in Rare Skin Disease Epidermolysis Bullosa. Skin pharmacology and physiology 9 33823521
2019 The Role of Marek's Disease Virus UL12 and UL29 in DNA Recombination and the Virus Lifecycle. Viruses 8 30696089
1999 Identification of the Marek's disease virus serotype 2 genes homologous to the glycoprotein B (UL27), ICP18.5 (UL28) and major DNA-binding protein (UL29) genes of herpes simplex virus type 1. The Journal of veterinary medical science 8 10563297
2012 Prokaryotic expression and characteristics of duck enteritis virus UL29 gene. Acta virologica 7 23237085
1986 Monoclonal antibodies, L-35 and L-36, define novel T cell activation antigens. Journal of immunology (Baltimore, Md. : 1950) 7 3095423
2014 Draft genome sequences and description of Lactobacillus rhamnosus strains L31, L34, and L35. Standards in genomic sciences 6 25197459
1996 Translational coupling in the Escherichia coli operon encoding translation initiation factor IF3 and ribosomal proteins L20 and L35. Biochimie 6 8955899
2024 TENT5A mediates the cancer-inhibiting effects of EGR1 by suppressing the protein stability of RPL35 in hepatocellular carcinoma. Cellular oncology (Dordrecht, Netherlands) 4 39570560
2021 Mycoplasma genitalium Protein of Adhesion Promotes the Early Proliferation of Human Urothelial Cells by Interacting with RPL35. Pathogens (Basel, Switzerland) 4 34832605
2018 Detection of Residual Proteins UL5 and UL29 Using a Targeted Proteomics Approach in HSV529, a Replication-Deficient HSV-2 Vaccine Candidate. Journal of pharmaceutical sciences 4 30176254
2013 Determination of HSV-1 UL5 and UL29 gene copy numbers in an HSV complementing Vero cell line. Journal of biotechnology 4 24140636
1996 Characterisation of genes encoding a nucleoside monophosphate kinase and a L35 ribosomal protein from Babesia bovis. Molecular and biochemical parasitology 4 8920009
2005 Ribosomal protein genes S23 and L35 from amphioxus Branchiostoma belcheri tsingtauense: identification and copy number. Acta biochimica et biophysica Sinica 3 16077906
2025 GADD45GIP1 promotes osteosarcoma progression by modulating RPL35 ubiquitination and alleviating endoplasmic reticulum stress via the PERK/eIF2α pathway. Cancer cell international 2 40604925
1996 The sequence of 23 kb surrounding the SNF3 locus on the left arm of yeast chromosome IV reveals the location of five known genes and characterizes at least six new open reading frames including putative genes for ribosomal protein L35 and a sugar transport protein. Yeast (Chichester, England) 2 8896272
2024 A Mutation in the Herpes Simplex Virus Type 1 (HSV-1) UL29 Gene is Associated with Anti-Herpesvirus Drugs' Susceptibility. Viruses 1 39772124
2026 Unresolved questions on the GADD45GIP1-RPL35 axis in osteosarcoma: mechanistic links to ER stress and therapeutic targeting. Cancer cell international 0 41606585
2025 Escherichia coli proteins uL29 and ACP stabilize the Tn7-encoded TnsD and its DNA binding. Mobile DNA 0 40847421
2025 Ribosomal protein L35 negatively regulates FMDV replication by recruiting AMFR to promote the ubiquitination and degradation of VP2. Journal of virology 0 41065387
2001 Nucleotide sequence of the porcine 60S ribosomal protein L35 cDNA. DNA sequence : the journal of DNA sequencing and mapping 0 11913794

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