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Cited by 223Genomics and Chromatin DynamicsAcute Myeloid Leukemia ResearchRetinoids in leukemia and cellular processes

Multiple Functional Domains of AML1: PU.1 and C/EBPα Synergize with Different Regions of AML1

Open access
Petrovick, Martha S., Hiebert, Scott W., Friedman, Alan D., Hetherington, Christopher J., Tenen, Daniel G., Zhang, Dong-Er·1998·Molecular and Cellular Biology
DOI 10.1128/mcb.18.7.3915PMC PMC108976
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Abstract

Control elements of many genes are regulated by multiple activators working in concert to confer the maximal level of expression, but the mechanism of such synergy is not completely understood. The promoter of the human macrophage colony-stimulating factor (M-CSF) receptor presents an excellent model with which we can study synergistic, tissue-specific activation for two reasons. First, myeloid-specific expression of the M-CSF receptor is regulated transcriptionally by three factors which are crucial for normal hematopoiesis: PU.1, AML1, and C/EBPalpha. Second, these proteins interact in such a way as to demonstrate at least two examples of synergistic activation. We have shown that AML1 and C/EBPalpha activate the M-CSF receptor promoter in a synergistic manner. As we report here, AML1 also synergizes, and interacts physically, with PU. 1. Detailed analysis of the physical and functional interaction of AML1 with PU.1 and C/EBPalpha has revealed that the proteins contact one another through their DNA-binding domains and that AML1 exhibits cooperative DNA binding with C/EBPalpha but not with PU.1. This difference in DNA-binding abilities may explain, in part, the differences observed in synergistic activation. Furthermore, the activation domains of all three factors are required for synergistic activation, and the region of AML1 required for synergy with PU.1 is distinct from that required for synergy with C/EBPalpha. These observations present the possibility that synergistic activation is mediated by secondary proteins contacted through the activation domains of AML1, C/EBPalpha, and PU.1.

Integrity signals

Registry

Crossref · 10.1128/mcb.18.7.3915

Cites

90 of 90 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

8 of 29 cited sources checked · 5 supported · 1 flagged · accuracy not yet measured

  • Study design and conductcitation

    Earlier Western blotting of transfected HepG2 extracts showed that these C/EBPα constructs produce protein.

    “The production of C/EBP␣ protein from these constructs has been demonstrated previously by Western blot analysis of extracts from transfected HepG2 cells (17).”
    Quote in the paperSubstantial claim1 of 1 sources checked
  • Premisecitation

    The region from amino acids 1 to 317 is important for synergy between AML1 and c-Myb.

    “a region important for synergy between AML1 and c-Myb (6)”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [6] source

      The Discussion (page unavailable) identifies AML-1B regions 216–289 and 278–480 as required for optimal cooperation with c-Myb, while saying its DNA-binding domain is dispensable. This supports the importance of part of amino acids 1–317, but not that the specified 1–317 region as a whole is the region required for synergy; the paper also identifies a required segment extending beyond residue 317. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • Findingown data

    A DNA-bound ternary complex formed with the AML1 runt domain and the C/EBPα bZip domain.

    “A ternary complex containing both proteins and the oligonucleotide was also observed (lane 4).”
    Quote in the paperSubstantial claim
  • Findingown data

    AML1 interacts physically with PU.1.

    “AML1 also synergizes, and interacts physically, with PU.1.”
    • Also stated in: § RESULTS · boosted
      “these data demonstrate that AML1 and PU.1 interact physically”
    Quote in the paperSubstantial claim
  • Findingown data

    Purified proteins allowed detection of both AML1-C/EBP and AML1-PU.1 DNA-protein ternary complexes.

    “these data demonstrate that AML1-C/EBP and AML1-PU.1 DNA-protein ternary complexes can be detected with purified proteins.”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 9
  • May combine two claims. 36
  • The quote only partly states this claim. 2

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    ERK-mediated phosphorylation of AML1A at serines 249 and 266 (AML1B serines 276 and 293) can raise the activity of an AML1-responsive reporter.

    “The activation of an AML1-responsive reporter by AML1A can be increased following phosphorylation by ERK on serines 249 and 266 (corresponding to serines 276 and 293 of AML1B) (68).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [68] source

      The Results (Fig. 2) identify the protein carrying the ERK-dependent sites as AML1b and state that AML1a lacks the PST region; no ERK-dependent phosphorylation shift was detected for AML1a. The Results (Fig. 3) assign the sites to AML1b serines 249 and 266 and say that its serine 276 was not a detectable ERK-dependent phosphorylation site. The reporter increase described in the Results (Fig. 8) therefore supports the effect for AML1b, not the isoform and residue mapping in the claim. (LLM, conf=0.91)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    CBP is a ubiquitous adapter protein that links transcription factors to the basal transcription machinery and is thought to stimulate transcription through contact with RNA polymerase II and histone acetylation.

    “CBP is a ubiquitous adapter protein that mediates contacts between transcription factors and the basal transcription machinery (1,26,33) and is thought to stimulate transcription both by physical contact with the RNA polymerase II complex and through intrinsic histone acetylation activity (4, 33, 52).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [1] source
    • cites [26] source
    • cites [33] source

      The discussion reports that CBP interacts with the basal transcription factor TFIIB, supporting the proposed link between transcription factors and basal machinery; the paper primarily investigates this bridging role in p300. Our copy does not establish that CBP is ubiquitous or that it stimulates transcription through contact with the RNA polymerase II complex or intrinsic histone acetylation. The indexed passages provide no page number. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Prior studies report that physical interaction between transcription factors can enhance DNA binding, which may explain synergy when two factors are present.

    “In several studies, the physical interaction between transcription factors increases DNA-binding ability, providing an explanation for synergistic activation in the presence of two factors (11,18,56,58,67).”
    Quote in the paperNot a substantial claim3 of 3 sources checked

  • May combine two claims.

    Premisecitation

    AML1 interacts with Ets-1, and in this case the runt domain contacts amino acids 123 to 240 in the Ets-1 N terminus rather than the conserved ets DNA-binding domain.

    “Although AML1 is known to interact with Ets-1, which like PU.1 is a member of the ets family of transcription factors, in this situation it is not the conserved ets DNA-binding domain which makes contact with the runt domain of AML1, but rather amino acids 123 to 240 in the N terminus of Ets-1 (18).”
    Quote in the paperNot a substantial claim1 of 1 sources checked

  • May combine two claims.

    Findingown data

    In HeLa cells, PU.1 alone gave 2-fold activation and AML1B alone 3.5-fold, but together they produced 12-fold activation.

    “Transfections of HeLa cells, which contain no endogenous PU.1 and little detectable AML1, showed that while PU.1 activates the promoter 2-fold and AML1B 3.5-fold, together they synergize to increase the activity of the promoter 12-fold (Fig. 5A).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: HeLa cells lack endogenous PU.1 and have only low detectable AML1.

  • May combine two claims.

    Findingown data

    Both purified GST-PU.1 and GST-runt bound a labeled probe carrying the AML1 and PU.1 sites from the M-CSF receptor promoter.

    “purified GST-PU.1 (lane 2) and GST-runt, a fusion protein containing the runt domain of AML1 (79) (lane 3), bound to the radiolabeled oligonucleotide which contains the M-CSF receptor promoter AML1 and PU.1 sites”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AML1B works with PU.1 to activate the M-CSF receptor promoter synergistically, using separate C-terminal regions for this and for its other synergy function.

    “Here we show that, in addition to C/EBP␣, AML1B interacts with PU.1 to synergistically activate the M-CSF receptor promoter but requires different regions contained within the C terminus for each function.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    GST-PU.1 pulled down both the 250- and 479-amino-acid AML1 forms, and the 479-amino-acid form appeared to bind more strongly.

    “Both the 250 (Fig. 4A, lanes 1 and 5)-and 479 (lanes 2 and 8)-amino-acid forms (AML1 and AML1B [Fig.1]) are pulled down by GST-PU.1;however, the interaction with AML1B appears stronger.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AML1 binds DNA cooperatively alongside C/EBPα, but not alongside PU.1.

    “AML1 exhibits cooperative DNA binding with C/EBP␣ but not with PU.1.”
    • Also stated in: § RESULTS · plain
      “while the runt domain of AML1 binds cooperatively to DNA in the presence of the C/EBP␣ bZip domain, the ternary complex with PU.1 is formed in a noncooperative manner.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The higher-order complex with the AML1 runt domain bound more DNA than GST-bZip alone, evidence that AML1 and C/EBPα bind DNA cooperatively.

    “we detected more DNA associated with the higher-order complex formed in the presence of the AML1 runt domain (lanes 10 to 18) than with GST-bZip (lanes 1 to 9), providing evidence that AML1 and C/EBP␣ exhibit cooperative DNA binding.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Removing PU.1's PEST domain did not change M-CSF receptor promoter activation or synergy with AML1B.

    “deletion of the PEST domain, which is important for interaction with NF-EM5 and activation of Bcell genes (58), has no effect on activation of the M-CSF receptor promoter or synergy with AML1B.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Less probe was shifted into the higher-order AML1-PU.1 complex than by GST-runt or GST-PU.1 alone, which indicates no cooperative DNA binding between AML1 and PU.1.

    “the amount of probe shifted by the higher-order complex is less than that shifted by either GST-runt (data not shown) or GST-PU.1,indicating that AML1 and PU.1 do not bind cooperatively to DNA.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    PU.1 lacking residues 1-70 or 33-100 lost synergy with AML1B, implicating residues 33-70 of its activation domain.

    “However, deletion of amino acids 1 to 70 or 33 to 100 abrogates synergy, indicating that amino acids 33 to 70 of the PU.1 activation domain are important for this function.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Full-length PU.1 and PU.1 residues 161-272, which include the ets domain, bound the GST-AML1 runt fusion specifically; PU.1 residues 1-161, the activation domain, did not.

    “full-length PU.1 (lanes 1 and 7) and amino acids 161 to 272 containing the ets domain of PU.1 (lanes 2 and 10), but not the activation domain of PU.1 (amino acids 1 to 161 [lanes 3 and 13]), interact specifically with the GST fusion protein containing the AML1 runt domain.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Ending AML1B at residue 381 left synergy intact, but ending it at residue 317 clearly disrupted synergy.

    “While termination of AML1B at amino acid 381 does not affect synergistic activation of the promoter, termination at amino acid 317 clearly does.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    PU.1-AML1B synergy is weaker than AML1B-C/EBPα synergy, yet it exceeds a purely additive effect.

    “Although the synergy between PU.1 and AML1B is weak relative to that observed with AML1B and C/EBP␣, it is more than an additive effect.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    No PU.1-AML1B synergy occurred when the PU.1 site, the AML1 site, or bp -86 to -37 was mutated or deleted, showing that DNA binding by both factors is needed.

    “However, this synergy is absent with reporter constructs bearing mutations in either the PU.1 or AML1 binding site [pM-CSF-R(mPU.1)-lucor pM-CSF-R(mAML1)luc, respectively] or a deletion of bp Ϫ86 to Ϫ37 [pM-CSF-R(DD)-luc], containing the binding sites for C/EBP, AML1, and PU.1, indicating that DNA binding is required for both factors.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: HeLa cells lack endogenous PU.1 and have only low detectable AML1.

  • May combine two claims.

    Findingown data

    AML1 and PU.1 bind each other physically, mainly via their DNA-binding domains.

    “these data demonstrate that AML1 and PU.1 interact physically and that this interaction occurs primarily through their DNA-binding domains.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In HeLa cells, AML1B(1-289) and AML1B(1-317) gave 3.5- and 3.8-fold promoter activation over pCMV5, and both synergized with C/EBPα.

    “in HeLa cells AML1B(1-289) and -(1-317) activate the M-CSF receptor promoter 3.5-and 3.8fold, respectively, relative to the expression vector, pCMV5, and both of these proteins are capable of synergizing with C/EBP␣ (Fig. 6A).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: HeLa cells lack endogenous PU.1 and have only low detectable AML1.

  • May combine two claims.

    Findingown data

    Unlabeled self probe, a PU.1-site probe, or an AML1-site probe competed for the PU.1-runt complex, whereas a nonspecific C/EBP-binding probe did not.

    “The formation of this PU.1-runtcomplex could be competed with a 200-fold molar excess of unlabeled self oligonucleotide (lane 5) or an oligonucleotide containing either a PU.1 binding site (lane 7) or an AML1 binding site (lane 8) but not with a nonspecific C/EBP-binding oligonucleotide (lane 6).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AML1B cut at residue 317 or 289 still synergized with C/EBPα, pointing to residues 268-317 as important for this activity.

    “AML1B truncated at either amino acid 317 or 289 retains the ability to synergize with C/EBP␣ (represented by the fold synergy), indicating that amino acids 268 to 317 are important for this activity.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Residues 268-317 of AML1B and the C/EBPα transactivation domains are essential for synergy.

    “The results of these experiments show that amino acids 268 to 317 of AML1B, and the transactivation domains of C/EBP␣, are critical for synergy.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AML1B-C/EBPα synergy depends on their transactivation domains, and the AML1 region needed for this synergy differs from the one needed for PU.1 synergy.

    “Synergy between AML1B and C/EBP␣ requires their transactivation domains, and the region of AML1 critical for this synergy is distinct from that important for synergy with PU.1.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Neither the 250-amino-acid AML1 isoform, ending at AML1B residue 268, nor AML1B truncated at residue 268 synergized with C/EBPα.

    “The 250-amino-acid isoform, AML1, which effectively terminates at residue 268 of AML1B, does not synergize with C/EBP␣, nor does AML1B truncated at amino acid 268.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AML1 and PU.1 synergistically activate the M-CSF receptor promoter, a function needing both DNA-binding sites and regions in the activation domains of AML1 and PU.1.

    “These data demonstrate that AML1 synergizes with PU.1 to activate the M-CSF receptor promoter and that this activity requires DNA-binding sites for both factors and regions contained within the activation domains of AML1 and PU.1.”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    AML1A and C/EBPα together activate the M-CSF receptor promoter synergistically.

    “Wild-type AML1A and mutant S249A/S266A both synergize with C/EBP␣.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    AML1 residues 1 to 317 are sufficient for synergy with C/EBPα but not with PU.1.

    “amino acids 1 to 317, a region important for synergy between AML1 and c-Myb (6), are sufficient for synergy with C/EBP␣ but not PU.1.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The 250-amino-acid AML1 form contains the runt domain but does not synergize with PU.1 or C/EBPα.

    “while the 250-amino-acid form of AML1 contains the runt domain, there is no synergy observed with either PU.1 or C/EBP␣.”
    Quote in the paperSubstantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    The runt domain and PU.1 did not show this increased binding; the ternary complex retained less probe than either PU.1 or the runt domain alone.

    “AML1 forms a tertiary complex on DNA with PU.1 but does not exhibit cooperative DNA binding.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    TBP binds a fusion protein containing the AML1B region needed for C/EBPα synergy but not a portion containing the domain needed for PU.1 synergy.

    “we are able to show that TBP binds to a fusion protein containing the region of AML1B important for synergy with C/EBP␣ but not to a portion that contains the domain required for synergy with PU.1 (data not shown).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Study design and conductown data

    Because the AML1B mutants activated the reporter, they are likely expressed, nuclear, and able to bind the promoter.

    “the activation of the reporter construct by the AML1B mutants indicates that they are expressed, are transported to the nucleus, and are capable of binding the promoter.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Study design and conductown data

    EMSA of COS extracts expressing the PU.1 constructs showed the mutant proteins were made at similar levels and could bind DNA.

    “EMSA analysis of COS cell extracts transfected with the PU.1 expression constructs demonstrated that the mutated proteins were produced in comparable amounts and are capable of binding to DNA (data not shown).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Physical interactions may help AML1 and C/EBPα bind DNA cooperatively, but they are not enough for strong synergy.

    “Therefore, while in the case of AML1 and C/EBP␣, the physical interactions may contribute to co- operative DNA binding, this is not sufficient for the strong synergy.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    The C-terminal domains of AML1B make specific, distinct contacts, and any of these contacts might contribute to synergistic activation.

    “confirming that the C-terminal domains of AML1B make specific and distinct contacts, any of which may play a role in synergistic activation.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    AML1's functional variability could be explained if it needs contacts with other factors to activate transcription and therefore depends on cell type and other DNA-binding sites.

    “The functional variability of AML1 can be explained if it requires contacts with other factors to activate transcription and is accordingly dependent on the cell type as well as other DNA-binding sites.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Kamei et al. proposed the term integrator for CBP, suggesting it integrates cellular signals that assemble transcription factors and coactivators and converts them into transcriptional activation.

    “Kamei et al. have coined the term “integrator,” postulating that CBP integrates diverse signals within the cell which culminate in the assembly of transcription factors and coactivators and translates them into transcriptional activation (26).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [26] source

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationnone given

    AML1's ability to synergize with PU.1 or C/EBPα, or to coordinate all three factors, might depend on coactivator and integrator availability or activation.

    “For instance, the ability for AML1 to synergize with PU.1 or C/EBP␣ or to coordinate the actions of all three transcription factors may depend on the activation or availability of coactivators and integrators.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Nuclear hormone receptors associate with steroid receptor coactivators and CBP, and their activation depends on efficient assembly of these complexes on DNA.

    “Recent reports have revealed a complex mechanism, whereby nuclear hormone receptors are associated with both steroid receptor coactivators and with CBP, and the activation ability of the transcription factor is dependent on the efficient assembly of these complexes on the DNA (43).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [43] source

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Prior work showed AML1B and C/EBPα synergistically activate the M-CSF receptor promoter, need both DNA-binding sites, and interact in vitro through their DNA-binding domains.

    “We have previously shown that AML1B and C/EBP␣ synergize to activate the M-CSF receptor promoter, that both DNAbinding sites are required for this function, and that the factors interact in vitro via their DNA-binding domains (runt and bZip domains, respectively) (79).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [79] source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    DNA-binding nuclear hormone receptors have been reported to use a comparable mechanism.

    “A similar mechanism has been described for the DNA-binding nuclear hormone receptors.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [43] source

  • May combine two claims.

    Interpretationown data

    The carboxy terminus of AML1B has at least two domains with different functions, one for synergy with C/EBPα and one for synergy with PU.1.

    “The carboxy terminus of AML1B contains at least two domains that serve disparate functions, each of which is responsible for synergy with C/EBP␣ and PU.1.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationown data

    Because the experiments used only the DNA-binding domains of AML1 and C/EBPα, the native proteins might behave differently.

    “However, since these experiments were performed with the DNA-binding domains of AML1 and C/EBP␣, we cannot exclude the possibility that the native pro-teins would exhibit different properties.”
    Quote in the paperNot a substantial claim
42 of 42

  • The paper relies on this without support.

    HeLa cells lack endogenous PU.1 and have only low detectable AML1.

    “Transfections of HeLa cells, which contain no endogenous PU.1 and little detectable AML1”
1 of 1

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • RUNX factors alone are relatively weak activators of transcription.

    “(RUNX factors alone are relatively weak activators of transcription.4,5,7,8,11)”

    From RUNX1 regulates corepressor interactions of PU.1 (2011) · cites this paper as [8]

    In the section on PU.1–AML1 synergy (page not available), AML1B alone activated the M-CSF receptor promoter 3.5-fold in HeLa cells, versus 12-fold with PU.1. This supports relatively modest activation by AML1B alone in that assay, but does not establish the claim for RUNX factors generally. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • RUNX1 physically interacts with PU.1, CEBPA, GATA1, GATA2, PAX5, FLI1, and ETS1.

    “RUNX1 physically interacts with PU.1, CEBPA, GATA1, GATA2, PAX5, FLI1, and ETS1.4–10,13”

    From RUNX1 regulates corepressor interactions of PU.1 (2011) · cites this paper as [8]

    The Results and Discussion (page unavailable in our copy) state that AML1/RUNX1 physically interacts with PU.1 and C/EBPα. The Discussion mentions cooperative DNA binding with Ets-1, but does not establish a physical interaction with it. The passages do not support the claimed physical interactions with GATA1, GATA2, PAX5, or FLI1. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • RUNX1 synergistically increases transcriptional activation by ETS1, PU.1(SPI1), CEBPA, GATA1, GATA2, and FLI1.

    “RUNX1 synergistically increases transcriptional activation by ETS1, PU.1(SPI1), CCAAT/enhancer binding protein-α (CEBPA), GATA1, GATA2, and FLI1.4–10”

    From RUNX1 regulates corepressor interactions of PU.1 (2011) · cites this paper as [8]

    The Results report synergistic activation of the M-CSF receptor promoter by AML1 (RUNX1) with PU.1, and the paper also reports synergy with C/EBPα. The Discussion describes cooperative binding with ETS1, but does not establish the claimed transcriptional synergy with ETS1; our copy provides no support for the GATA1, GATA2, or FLI1 parts of the list. Page numbers were not available in the indexed passages. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • RUNX1 cooperates with lineage-specifying TFs (e.g., PU.1/SPI1) to activate myeloid differentiation genes such as MCSFR and GMCSFR.

    “The transcription factor (TF) RUNX1 cooperates with lineage-specifying TFs (eg, PU.1/SPI1) to activate myeloid differentiation genes, such as macrophage and granulocyte macrophage colony-stimulating factor receptors (MCSFR and GMCSFR).”

    From RUNX1 regulates corepressor interactions of PU.1 (2011) · cites this paper as [8]

    The section on PU.1–AML1 synergy reports that AML1B (RUNX1) and PU.1 together activate the M-CSF receptor promoter more than additively. The paper does not show their cooperation in activating the GM-CSF receptor gene; its mention of GM-CSF concerns the cytokine gene, not its receptor. The cited result also concerns promoter activity in transfected cells rather than direct measurement of differentiation. (LLM, conf=0.89)

    Experimental — accuracy not yet measured

  • In cotransfection experiments, RUNX1, PU.1, and CEBPA play essential roles in activation of the hematopoietic differentiation genes MCSFR and GMCSFR.

    “In cotransfection experiments, RUNX1, PU.1, and CEBPA play essential roles in activation of the hematopoietic differentiation genes MCSFR and GMCSFR.7,8,31”

    From RUNX1 regulates corepressor interactions of PU.1 (2011) · cites this paper as [8]

    The paper’s Abstract and its section on PU.1–AML1 synergy report transient-transfection evidence that AML1 (RUNX1), PU.1, and C/EBPα contribute to activation of the M-CSF receptor promoter. The cited paper does not show the three factors activating the GM-CSF receptor gene; its References instead list a separate study of PU.1 and C/EBPα regulation of that gene. Page numbers are unavailable in our copy. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Aligned AML1 regions differ by one residue in length

    Minor

    The two regions described as identical contain different numbers of residues when both endpoints are included: AML1 residues 5-242 span 238 residues, while AML1B residues 32-268 span 237.

  • PU.1 mutant has two reported endpoints

    Minor

    The Results describe the PU.1 activation-domain construct in Figure 4C as ending at amino acid 161, but the figure caption identifies that construct as ending at amino acid 163.

2 of 2

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Related

Authors
Petrovick, M. S., Hiebert, S. W., Friedman, A. D., Hetherington, C. J., Tenen, D. G., & Zhang, D.-E.
Affiliations
  • Beth Israel Deaconess Medical Center (US)
  • Vanderbilt University (US)
  • Johns Hopkins University (US)
ORCID
Venue
Molecular and Cellular Biology
Publisher
Informa UK Limited
Published
1998-07
Volume
18
Issue
7
Pages
3915-3925
ISSN
  • 0270-7306 (ISSN-L)
  • 1098-5549
DOI
10.1128/mcb.18.7.3915
PMC
PMC108976
OpenAlex
W2138483214
Cited by
223
References
90
Citations by year
2026: 1 · 2025: 1 · 2024: 2 · 2023: 3 · 2022: 3 · 2021: 3 · 2020: 3 · 2019: 5 · 2018: 6 · 2017: 7 · 2016: 3 · 2015: 3 · 2014: 4 · 2013: 4 · 2012: 5
FWCI
6.03
Type
journal article
Language
en
Open access
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Molecular and Cellular Biology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Health Sciences › Medicine › Hematology › Acute Myeloid Leukemia Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinoids in leukemia and cellular processes
Keywords
RUNX1, PU.1, C/EBPα, transcriptional synergy, cooperative DNA binding, transcription factor interaction, transactivation domain, hematopoiesis
MeSH terms
Proto-Oncogene Protein Spi-1, Animals, Binding Sites, Cell Line

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-07.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-07.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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