Functional Annotation Report: *Saccharomyces cerevisiae* SWI4 Falcon Edison Scientific Literature 26 citations 1 artifacts 2026-09-25T04:16:01.904528

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Functional Annotation Report: Saccharomyces cerevisiae SWI4

Executive summary

The requested protein is correctly identified as Swi4, encoded by SWI4/YER111C in Saccharomyces cerevisiae strain S288c and represented by UniProt P25302. Organism-specific literature identifies Swi4 as the sequence-specific DNA-binding subunit of SBF—the Swi4/Swi6 cell-cycle transcription factor—and not as an enzyme, transporter, or structural cell-wall protein. No evidence from similarly named proteins in other organisms was used to annotate this target. The supplied ankyrin-repeat, APSES/KilA-like helix-turn-helix, and SWI6-like annotations agree with experimentally mapped Swi4 regions. (baetz1999regulationofcell pages 1-2, su2024controlofmeiotic pages 2-4, siegmund1996thesaccharomycescerevisiae pages 1-2)

Swi4’s primary function is to recognize SCB promoter elements, with the canonical consensus 5′-CACGAAA-3′, and recruit/regulate transcriptional machinery with Swi6. SBF activates a late-G1 transcriptional program that couples commitment at Start to G1-cyclin production, bud emergence, cell-wall synthesis, and other events required for a new division cycle. Swi4 acts primarily in the nucleus, where its activity is controlled by Swi6, Whi5, G1 and mitotic cyclin–Cdk1 complexes, promoter occupancy, and cell-state-specific regulation. (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2, nishikawa2008asystematiccell pages 126-133)

1. Identity verification and nomenclature

The literature specifically identifies budding-yeast SWI4/YER111C as the gene encoding the Swi4 component of SBF; SBF contains Swi4 plus Swi6, whereas the related MBF complex substitutes Mbp1 for Swi4 while retaining Swi6. This establishes that the target is the S288c budding-yeast protein described in the query rather than a similarly named protein from another fungus. (su2024controlofmeiotic pages 2-4)

The alternative name ART1 should not be confused with unrelated proteins carrying ART-family terminology. For functional annotation, SWI4/YER111C/P25302 is the unambiguous identifier set.

2. Primary molecular function

2.1 Sequence-specific transcription factor

Swi4 is the DNA-recognition and promoter-targeting subunit of SBF. SBF binds repeated SCB elements in promoters and activates transcription around the G1/S transition. The best-supported SCB consensus is CACGAAA. Swi6 is principally regulatory: it interacts with Swi4, relieves Swi4 autoinhibition, and participates in cell-cycle-dependent control of transcription. Thus, the closest substrate-specificity analogy is recognition of SCB-containing double-stranded promoter DNA—not catalysis of a chemical reaction. (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)

Canonical experimentally supported SBF-responsive genes include CLN1, CLN2, PCL1, PCL2, HO, and multiple genes required for budding or cell-wall biosynthesis. CLN1 and CLN2 encode G1 cyclins, creating positive feedback in which SBF-dependent cyclin expression further increases G1-Cdk1 activity and reinforces Start. HO links SBF regulation to mating-type switching, while morphogenesis and wall-biosynthesis targets coordinate physical bud formation with cell-cycle commitment. (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)

2.2 Molecular architecture and mechanism

Experimental dissection supports three principal functional regions:

  1. N-terminal DNA-binding region. The first approximately 197 residues contain the sequence-specific DNA-binding activity. This agrees with the supplied KilA/APSES helix-turn-helix annotations.
  2. Central ankyrin-repeat region. Four ankyrin repeats mediate protein interactions, including association with mitotic Clb2–Cdc28/Cdk1.
  3. Conserved C-terminal region. This region binds Swi6 and also participates in intramolecular autoinhibition of the N-terminal DNA-binding domain. (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)

Full-length Swi4 is poorly competent for SCB binding without Swi6 because its C terminus masks the N-terminal DNA-binding region. Mutations in or removal of the extreme C terminus restore DNA binding or transcriptional activity, and direct interaction between C- and N-terminal fragments supports a physical autoinhibitory mechanism. Evidence that full-length Swi4 is monomeric favors intramolecular masking rather than inhibition through Swi4 oligomerization. Swi6 binding is therefore not merely complex assembly: it changes Swi4 into a DNA-binding-competent regulatory state. (baetz1999regulationofcell pages 1-2)

The supplied domain list is consequently coherent with the literature: the APSES/KilA-like region explains DNA recognition, ankyrin repeats provide interaction surfaces, and the SWI6-like/conserved C-terminal region supports regulated partner binding.

3. Cellular localization

Swi4 is predominantly nuclear and was reported to remain nuclear throughout the mitotic cell cycle. Its activity is therefore not regulated mainly by complete Swi4 nuclear export. Instead, regulation occurs through its autoinhibited versus Swi6-bound state, recruitment of Whi5, cyclin–Cdk1-dependent phosphorylation, changing promoter occupancy, and the cell-cycle-dependent localization of Swi6. Swi6 is enriched in the nucleus during late mitosis/G1 but becomes more cytoplasmic during S, G2, and early mitosis. (baetz1999regulationofcell pages 1-2)

The physiologically relevant site of Swi4 action is consequently chromatin in the nucleus, especially SCB-containing promoters. There is no credible evidence that Swi4 is secreted, membrane-associated, or a structural constituent of the cell wall.

4. Pathway placement and regulatory cycle

4.1 Pre-Start repression

Before Start, SBF can occupy promoters in a transcriptionally restrained complex containing Whi5. Whi5 recruitment to CLN2 and PCL1 promoters depends on SWI4, supporting direct repression through promoter-bound SBF. Following release from G1 arrest, Whi5 disappeared from the PCL1 promoter within approximately 30 minutes, while Swi6 remained transiently associated. Deleting WHI5 advances G1/S transcription to a smaller mean cell size but does not abolish its periodicity; Whi5 therefore controls onset more strongly than the basic oscillator. (nishikawa2008asystematiccell pages 126-133)

4.2 Activation at Start

As cells grow, G1 cyclin–Cdk1 activity rises. Cln3/Cdk1 and downstream Cln1/2-Cdk1-associated events relieve repression, permitting SBF to activate CLN1/2 and the broader budding/morphogenesis program. Older models emphasized direct Whi5 phosphorylation; current synthesis recognizes that phosphorylation of Whi5 or Swi6 can provide partially redundant routes to timely SBF activation. A double non-phosphorylatable WHI5-12A SWI6-SA4 mutant was reported to be approximately 40% larger, indicating substantial functional overlap rather than a single obligatory phosphorylation switch. (ravi2024modelingthestart pages 5-6)

4.3 Shutoff after G1/S

SBF must be turned off after its G1 pulse. Mitotic Clb–Cdc28/Cdk1 complexes repress SBF, and the Swi4 ankyrin repeats physically associate with Clb2–Cdc28 in vivo. Later cyclin-dependent regulation promotes loss of SBF from promoters during G2/M, helping ensure that G1 cyclins are activated once rather than repeatedly in the same cycle. (siegmund1996thesaccharomycescerevisiae pages 1-2, ravi2024modelingthestart pages 26-28)

This architecture separates two regulatory operations: G1 cyclin–Cdk1 activates a previously restrained promoter complex, while mitotic cyclin–Cdk1 contributes to promoter dissociation and termination of the transcriptional pulse.

5. Biological processes governed by Swi4

Start, budding, and cell-wall construction

Swi4’s central biological role is to coordinate Start-associated transcription with the cellular structures needed for division. SBF targets include G1 cyclins and genes involved in budding and cell-wall biosynthesis. Swi4 therefore regulates wall construction indirectly through transcription; it is not itself a wall enzyme or scaffold. (baetz1999regulationofcell pages 1-2)

Mating-type switching

SBF regulation of HO, together with cell-cycle-dependent control at relevant loci, links Swi4 to the timing of mating-type switching. This is a specialized extension of its nuclear transcription-factor function rather than a separate catalytic activity. (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)

Mitosis-to-meiosis transition

Recent work shows that Swi4 must be actively suppressed when cells enter meiosis. Swi4 abundance declined by approximately 30% after two hours of meiotic induction, whereas Mbp1 and Swi6 increased. A long undecoded transcript isoform, SWI4 LUTI, represses the downstream canonical SWI4 promoter and reduces translation through upstream open reading frames; this mechanism cooperates with Whi5 to restrict SBF. (su2024controlofmeiotic pages 2-4)

Forced SWI4 expression prematurely activated SBF targets—including CLN1, CLN2, and PCL1—reduced early meiotic gene expression, and delayed meiotic entry. Deleting CLN1 or CLN2 partially rescued that delay. The mechanistic interpretation is that excess G1 cyclins disrupt interaction of the meiotic regulator Ime1 with Ume6, thereby antagonizing the meiotic program. Reported statistical effects included CLN1 p=0.0351, CLN2 p=0.0013, and altered meiotic progression with Mann–Whitney p=0.0045. (su2024controlofmeiotic pages 16-18, su2024controlofmeiotic pages 2-4)

6. Recent developments, 2023–2024

6.1 Dual restriction of SBF during meiotic entry

Su, Yendluri, and Ünal’s work, available in its cited February 2024 version, extends Swi4 biology beyond the canonical mitotic Start switch. It establishes that transcriptional interference/translation control through SWI4 LUTI and protein-level repression through Whi5 jointly prevent inappropriate activation of the mitotic SBF program during meiotic entry. This is direct experimental evidence that Swi4 abundance itself is a regulated determinant of cell-state choice. Publication: Control of meiotic entry by dual inhibition of a key mitotic transcription factor, DOI: https://doi.org/10.7554/eLife.90425.2. (su2024controlofmeiotic pages 16-18, su2024controlofmeiotic pages 2-4)

6.2 START-BYCC mechanistic model

Ravi, Samart, and Zwolak published START-BYCC in PLOS Computational Biology on August 2, 2024. The model explicitly represents Swi4/SBF, Swi6, Whi5, Mbp1/MBF, cyclin-dependent phosphorylation, promoter binding, and nuclear–cytoplasmic transport. It expanded the Start subsystem from one species and eight parameters in an earlier model to 51 species and 56 parameters. (ravi2024modelingthestart pages 25-26, ravi2024modelingthestart pages 6-9)

The authors simulated more than 200 mutants, including over 100 Start mutants, and reported successful fitting of approximately 95% of mutant phenotypes. The model reproduced viability, cell-size, Start-timing, and rescue relationships; examples included rescue of cln3Δ swi4Δ by whi5Δ, sic1Δ, or GAL-BCK2, and rescue of swi4Δ swi6Δ by GAL-CLB5 or GAL-CLN2 but not GAL-CLN3. Publication: Modeling the START transition in the budding yeast cell cycle, DOI: https://doi.org/10.1371/journal.pcbi.1012048. (ravi2024modelingthestart pages 23-25, ravi2024modelingthestart pages 9-10)

These results are valuable as quantitative integration, but they are not independent biochemical proof for every modeled interaction. Assumptions include compartment-volume ratios, Hill-function approximations for multisite phosphorylation, inferred stoichiometries, and lumped cyclin species. The model should therefore guide perturbation design and consistency checking rather than replace direct promoter-occupancy, biochemical, or live-cell experiments. (ravi2024modelingthestart pages 26-28, ravi2024modelingthestart pages 25-26)

7. Evidence map

The following table distinguishes direct experimental findings from computational synthesis.

Feature/question Best-supported annotation Evidence type Key source with year and DOI/URL
Identity SWI4/YER111C encodes Swi4, the S. cerevisiae SBF-specific DNA-binding subunit; this matches UniProt P25302 and is distinct from similarly named proteins in other fungi. Curated identity plus organism-specific experimental literature Su et al. (2024), eLife 90425.2 (su2024controlofmeiotic pages 2-4)
Primary molecular function / SBF Swi4 forms SBF (SCB-binding factor) with the regulatory subunit Swi6. Swi4 supplies sequence-specific promoter recognition; Swi6 relieves Swi4 autoinhibition and supports regulated transcription. It is a transcription factor, not an enzyme or transporter. Direct DNA-binding, protein-interaction, and mutational experiments Baetz & Andrews (1999), 10.1128/MCB.19.10.6729 (baetz1999regulationofcell pages 1-2)
DNA motif / substrate specificity SBF recognizes repeated SCB elements, consensus 5′-CACGAAA-3′, in target promoters. The molecular substrate is promoter DNA rather than a small molecule. Direct biochemical DNA-binding and promoter-footprinting evidence Siegmund & Nasmyth (1996), 10.1128/MCB.16.6.2647 (siegmund1996thesaccharomycescerevisiae pages 1-2)
Domains and architecture The N-terminal region contains the APSES/KilA-like DNA-binding domain; the first 197 residues are sufficient for DNA binding. A central region contains four ankyrin repeats, which associate with Clb2–Cdc28, while the conserved C terminus binds Swi6 and intramolecularly masks the DNA-binding region. This agrees with the supplied InterPro APSES, ankyrin-repeat, and SWI6-like annotations. Domain deletions, point mutants, binding assays, monomer-state analysis, and co-immunoprecipitation Baetz & Andrews (1999), 10.1128/MCB.19.10.6729; Siegmund & Nasmyth (1996), 10.1128/MCB.16.6.2647 (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)
Cellular localization Swi4 is principally nuclear and remains nuclear throughout the mitotic cell cycle. Regulation depends substantially on promoter occupancy, Swi6 localization, and inhibitory/activating partners rather than wholesale Swi4 nuclear export. Direct protein-localization experiments Baetz & Andrews (1999), 10.1128/MCB.19.10.6729 (baetz1999regulationofcell pages 1-2)
Canonical targets and processes Well-supported targets include CLN1, CLN2, PCL1, PCL2, HO, and genes involved in budding and cell-wall biosynthesis. Accordingly, Swi4 coordinates G1/S cyclin production, bud emergence, morphogenesis, mating-type switching, and Start-associated transcription. Promoter analysis, genetics, expression assays, and DNA occupancy Baetz & Andrews (1999), 10.1128/MCB.19.10.6729; Siegmund & Nasmyth (1996), 10.1128/MCB.16.6.2647 (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)
Start activation Before Start, promoter-bound SBF is restrained by Whi5. Rising G1 cyclin–Cdk1 activity promotes release/inactivation of repression and SBF-dependent positive feedback through CLN1/2. Whi5 occupancy at CLN2/PCL1 is Swi4-dependent and disappears from PCL1 within approximately 30 min after release from G1 arrest. Direct ChIP/promoter-occupancy, genetics, and cell-cycle experiments Nishikawa (2008), summarized evidence on Swi4-dependent Whi5 recruitment (nishikawa2008asystematiccell pages 126-133); Ravi et al. (2024), 10.1371/journal.pcbi.1012048 (ravi2024modelingthestart pages 5-6)
G2 shutoff Mitotic Clb–Cdc28 activity represses SBF: Swi4 ankyrin repeats bind Clb2–Cdc28, and phosphorylation-associated regulation promotes loss of SBF from promoters in G2/M, preventing repeated G1 transcription within one cycle. Direct co-immunoprecipitation and cell-cycle promoter-occupancy evidence; later computational integration Siegmund & Nasmyth (1996), 10.1128/MCB.16.6.2647; Ravi et al. (2024), 10.1371/journal.pcbi.1012048 (siegmund1996thesaccharomycescerevisiae pages 1-2, ravi2024modelingthestart pages 26-28)
Cell-wall role SBF directly links cell-cycle entry to expression of cell-wall biosynthetic and budding genes. This is a transcriptional coordination role rather than a structural cell-wall function; stress-responsive signaling can additionally act through the Swi4/Swi6 system. Direct target-gene and genetic evidence; pathway interpretation Baetz & Andrews (1999), 10.1128/MCB.19.10.6729 (baetz1999regulationofcell pages 1-2)
2024 meiosis development During meiotic entry, Swi4 abundance falls by approximately 30% after 2 h. A SWI4 LUTI transcript and Whi5 jointly suppress SBF; inappropriate SWI4 expression activates CLN1/CLN2/PCL1, reduces early-meiotic transcription, and delays meiosis. Reported effects included CLN1 p=0.0351, CLN2 p=0.0013, and meiotic progression p=0.0045. Direct time-course, expression, genetic, and meiotic-progression experiments Su et al. (2024 version), 10.7554/eLife.90425.2 (su2024controlofmeiotic pages 16-18, su2024controlofmeiotic pages 2-4)
2024 START-BYCC model START-BYCC explicitly represents Swi4/SBF, Swi6, Whi5, phosphorylation, promoter binding, and nuclear–cytoplasmic transport. It expanded the START module from 1 species/8 parameters to 51 species/56 parameters, simulated more than 200 mutants including over 100 START mutants, and reported approximately 95% successful phenotype fitting. These values reflect computational synthesis and validation against published phenotypes—not new direct biochemical measurements of Swi4. Computational mechanistic synthesis using nonlinear ODEs, mutant-phenotype fitting, and independent numerical checks Ravi, Samart & Zwolak (published August 2, 2024), 10.1371/journal.pcbi.1012048 (ravi2024modelingthestart pages 9-10, ravi2024modelingthestart pages 6-9)

Table: Compact evidence map for the verified S. cerevisiae Swi4 protein P25302/YER111C, separating direct experimental findings from the 2024 START-BYCC computational synthesis.

8. Current applications and real-world implementations

Swi4 is primarily a research-system regulator, not a clinical target with an approved therapeutic application. Its established applications include:

9. Expert assessment and remaining uncertainties

The strongest functional annotation is: nuclear, sequence-specific DNA-binding transcription factor; SBF-specific subunit controlling the G1/S transcriptional program. This conclusion is supported by direct DNA-binding studies, domain mutations, protein-interaction assays, promoter occupancy, localization, and classical genetics. The autoinhibition/Swi6-release mechanism and SCB specificity are especially well supported. (baetz1999regulationofcell pages 1-2, siegmund1996thesaccharomycescerevisiae pages 1-2)

Several qualifications are important:

  1. SBF output is context-dependent. Promoter sequence, chromatin, Swi6, Whi5, Cdk1 activities, and cell state all shape transcription; an SCB motif alone is not sufficient to infer physiological regulation.
  2. Localization and activity are distinct. Swi4 can remain nuclear while SBF activity changes sharply, so nuclear detection should not be equated with active promoter transcription. (baetz1999regulationofcell pages 1-2)
  3. Start is not a one-substrate phosphorylation switch. Current evidence supports partially redundant regulation through Whi5, Swi6, Bck2, and cyclin-CDK activities. (ravi2024modelingthestart pages 5-6, ravi2024modelingthestart pages 6-9)
  4. Recent quantitative models are integrative, not definitive. High mutant-fit rates demonstrate internal explanatory power, but fitted parameters and lumped reactions need experimental validation. (ravi2024modelingthestart pages 25-26, ravi2024modelingthestart pages 9-10)
  5. Recent mechanistic expansion is strongest for meiosis. The 2024 LUTI study adds direct evidence for regulation of SWI4 expression itself and for antagonism between mitotic SBF output and meiotic entry. (su2024controlofmeiotic pages 16-18, su2024controlofmeiotic pages 2-4)

Conclusion

SWI4/YER111C encodes the nuclear transcription factor Swi4, UniProt P25302. Through its N-terminal APSES-like DNA-binding domain, Swi4 recognizes SCB promoter elements; its ankyrin repeats and conserved C terminus integrate interactions with cyclin–Cdk1 complexes and Swi6. In SBF, it activates G1 cyclins, budding, and cell-wall-biogenesis genes at Start. Whi5 restrains promoter-bound SBF before commitment, G1 cyclin–Cdk1 activity promotes activation, and mitotic Clb–Cdk1 activity contributes to shutoff. Recent research has broadened this canonical annotation by showing that SWI4 expression is actively repressed during meiotic entry and by incorporating Swi4 regulation into a quantitatively validated whole-Start model. (baetz1999regulationofcell pages 1-2, su2024controlofmeiotic pages 16-18, siegmund1996thesaccharomycescerevisiae pages 1-2, ravi2024modelingthestart pages 9-10)

References

  1. (baetz1999regulationofcell pages 1-2): Kristin Baetz and Brenda Andrews. Regulation of cell cycle transcription factor swi4 through auto-inhibition of dna binding. Molecular and Cellular Biology, 19:6729-6741, Oct 1999. URL: https://doi.org/10.1128/mcb.19.10.6729, doi:10.1128/mcb.19.10.6729. This article has 63 citations and is from a domain leading peer-reviewed journal.

  2. (su2024controlofmeiotic pages 2-4): Amanda J. Su, Siri C. Yendluri, and Elçin Ünal. Control of meiotic entry by dual inhibition of a key mitotic transcription factor. ArXiv, Feb 2024. URL: https://doi.org/10.7554/elife.90425.2, doi:10.7554/elife.90425.2. This article has 11 citations.

  3. (siegmund1996thesaccharomycescerevisiae pages 1-2): Robert F. Siegmund and Kim A. Nasmyth. The saccharomyces cerevisiae start-specific transcription factor swi4 interacts through the ankyrin repeats with the mitotic clb2/cdc28 kinase and through its conserved carboxy terminus with swi6. Molecular and Cellular Biology, 16:2647-2655, Jun 1996. URL: https://doi.org/10.1128/mcb.16.6.2647, doi:10.1128/mcb.16.6.2647. This article has 108 citations and is from a domain leading peer-reviewed journal.

  4. (nishikawa2008asystematiccell pages 126-133): JL Nishikawa. A systematic cell size screen in saccharomyces cerevisiae and characterization of the start inhibitor whi5. Unknown journal, 2008.

  5. (ravi2024modelingthestart pages 5-6): Janani Ravi, Kewalin Samart, and Jason Zwolak. Modeling the start transition in the budding yeast cell cycle. Aug 2024. URL: https://doi.org/10.1371/journal.pcbi.1012048, doi:10.1371/journal.pcbi.1012048. This article has 1 citations and is from a highest quality peer-reviewed journal.

  6. (ravi2024modelingthestart pages 26-28): Janani Ravi, Kewalin Samart, and Jason Zwolak. Modeling the start transition in the budding yeast cell cycle. Aug 2024. URL: https://doi.org/10.1371/journal.pcbi.1012048, doi:10.1371/journal.pcbi.1012048. This article has 1 citations and is from a highest quality peer-reviewed journal.

  7. (su2024controlofmeiotic pages 16-18): Amanda J. Su, Siri C. Yendluri, and Elçin Ünal. Control of meiotic entry by dual inhibition of a key mitotic transcription factor. ArXiv, Feb 2024. URL: https://doi.org/10.7554/elife.90425.2, doi:10.7554/elife.90425.2. This article has 11 citations.

  8. (ravi2024modelingthestart pages 25-26): Janani Ravi, Kewalin Samart, and Jason Zwolak. Modeling the start transition in the budding yeast cell cycle. Aug 2024. URL: https://doi.org/10.1371/journal.pcbi.1012048, doi:10.1371/journal.pcbi.1012048. This article has 1 citations and is from a highest quality peer-reviewed journal.

  9. (ravi2024modelingthestart pages 6-9): Janani Ravi, Kewalin Samart, and Jason Zwolak. Modeling the start transition in the budding yeast cell cycle. Aug 2024. URL: https://doi.org/10.1371/journal.pcbi.1012048, doi:10.1371/journal.pcbi.1012048. This article has 1 citations and is from a highest quality peer-reviewed journal.

  10. (ravi2024modelingthestart pages 23-25): Janani Ravi, Kewalin Samart, and Jason Zwolak. Modeling the start transition in the budding yeast cell cycle. Aug 2024. URL: https://doi.org/10.1371/journal.pcbi.1012048, doi:10.1371/journal.pcbi.1012048. This article has 1 citations and is from a highest quality peer-reviewed journal.

  11. (ravi2024modelingthestart pages 9-10): Janani Ravi, Kewalin Samart, and Jason Zwolak. Modeling the start transition in the budding yeast cell cycle. Aug 2024. URL: https://doi.org/10.1371/journal.pcbi.1012048, doi:10.1371/journal.pcbi.1012048. This article has 1 citations and is from a highest quality peer-reviewed journal.

Artifacts

Citations

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