Affinage

SMAD4

SMAD family member 4 · UniProt Q13485

Length
552 aa
Mass
60.4 kDa
Annotated
2026-06-10
100 papers in source corpus 32 papers cited in narrative 32 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

SMAD4 (DPC4) is the central co-SMAD that transduces TGF-β superfamily signals into transcriptional responses and acts as a tumor suppressor first identified through homozygous deletion and intragenic mutation at chromosome 18q21.1 in pancreatic carcinoma (PMID:8553070). Upon ligand stimulation it forms regulated heteromeric complexes with receptor-activated R-SMADs — Smad1 in response to BMP and Smad2 in response to activin/TGF-β — interactions that drive mesoderm induction and antimitogenic responses (PMID:8893010). Within these complexes SMAD4 contributes two separable functions: its N-terminal MH1 domain promotes DNA binding while its C-terminal MH2 domain supplies a transcriptional activation function, with the MH2 domain also mediating the homo- and heteromeric associations that cancer-associated mutations disrupt (PMID:9389648, PMID:9111321, PMID:10871368). SMAD4 stabilizes DNA-binding transcription-factor complexes (e.g., the Smad2–FAST-1 ARF complex) and recruits the coactivator CBP/p300 to activate TGF-β-responsive transcription (PMID:9288972, PMID:9679060). Genetically it is essential for early development, as Smad4-null mice fail to gastrulate and form mesoderm owing to defective visceral endoderm and epiblast proliferation (PMID:9420335, PMID:9520423). SMAD4 activity is tuned by a dense layer of post-translational control: MAPK/GSK3-primed linker phosphorylation generates a β-TrCP phosphodegron targeting SMAD4 for SCF(β-TrCP1)-mediated proteasomal degradation (PMID:14988407, PMID:25373906); SMURF2-mediated inhibitory monoubiquitination is reversed by the deubiquitinases USP4 and USP10 (PMID:28468752, PMID:31721429); Wip1 dephosphorylates Thr277 to limit nuclear accumulation (PMID:32103600); PRMT5-mediated R361 methylation is required for SMAD complex assembly and nuclear import (PMID:36991117); ALK-mediated Tyr95 phosphorylation blocks DNA binding (PMID:30664791); and SFPQ sequesters SMAD4 in phase-separated condensates to exclude it from chromatin (PMID:38103553). Beyond canonical transcription, SMAD4 enforces tissue identity and suppresses tumor progression — shifting the angiogenic balance by repressing VEGF and inducing thrombospondin-1 (PMID:10944227), restraining endothelial PI3K/AKT signaling through CK2 to prevent arteriovenous malformations (PMID:29976569), stabilizing enterocyte identity via a reinforcing feed-forward loop with HNF4 (PMID:30988513), and suppressing Wnt/β-catenin-driven dedifferentiation and metastatic effectors such as FOSL1 (PMID:14597578, PMID:34320363). Germline/somatic loss in epithelial contexts thus de-represses pro-tumorigenic programs, with DPC4 inactivation occurring late in pancreatic neoplastic progression (PMID:10766191).

Mechanistic history

Synthesis pass · year-by-year structured walk · 26 steps
  1. 1996 High

    Established SMAD4/DPC4 as a candidate tumor suppressor and placed it in a TGF-β-like pathway, answering where the gene sits in cancer genetics.

    Evidence Homozygous deletion mapping and sequencing of pancreatic tumor DNA at 18q21.1

    PMID:8553070

    Open questions at the time
    • Did not define the biochemical activity of the protein
    • Pathway link inferred from sequence similarity to Mad rather than direct biochemistry
  2. 1996 High

    Defined SMAD4 as a ligand-regulated partner of multiple R-SMADs, answering how it couples to distinct TGF-β superfamily ligands.

    Evidence Co-immunoprecipitation plus Xenopus and mammalian functional assays

    PMID:8893010

    Open questions at the time
    • Did not map the domains mediating each interaction
    • Did not establish nuclear vs cytoplasmic site of complex assembly
  3. 1997 High

    Resolved the molecular logic of SMAD4 within transcriptional complexes — MH1 for DNA binding, MH2 for activation and oligomerization — explaining why cancer mutations cluster in MH2.

    Evidence Domain deletion/chimera constructs, yeast two-hybrid, ARF complex reconstitution, reporter assays

    PMID:9111321 PMID:9153220 PMID:9288972 PMID:9389648

    Open questions at the time
    • Functional contribution of the linker region characterized in only a single study
    • Did not provide atomic structure of the complexes
  4. 1997 Medium

    Demonstrated that SMAD4 re-expression restores TGF-β growth inhibition in SMAD4-null tumor cells, establishing causal tumor-suppressive function.

    Evidence Reconstitution in MDA-MB-468 cells with growth inhibition and reporter readouts

    PMID:9150356

    Open questions at the time
    • Single cell line
    • Did not separate growth-arrest from later angiogenic functions
  5. 1998 High

    Showed SMAD4 is essential for gastrulation and mesoderm formation, defining its non-redundant developmental requirement.

    Evidence Two independent knockout mouse lines with tetraploid rescue and blastocyst outgrowth analysis

    PMID:9420335 PMID:9520423

    Open questions at the time
    • Lethality precludes analysis of later tissue-specific roles
    • Did not identify the downstream developmental targets
  6. 1998 High

    Identified CBP/p300 as the coactivator SMAD4 recruits, answering how the complex drives transcription.

    Evidence Co-IP, reporter assays, and E1A inhibition

    PMID:9679060

    Open questions at the time
    • Did not map promoter-specific recruitment
    • Did not address coactivator competition in vivo
  7. 2000 High

    Revealed a transcription-independent tumor-suppressive output — angiogenesis suppression via VEGF down and thrombospondin-1 up — broadening SMAD4 function beyond growth arrest.

    Evidence Stable reconstitution in pancreatic cancer cells with in vivo nude-mouse tumor and expression analysis

    PMID:10944227

    Open questions at the time
    • Mechanism of VEGF/TSP-1 regulation not resolved at promoter level
    • Restoration occurred without restoring TGF-β sensitivity
  8. 2000 High

    Mapped MH1 DNA-binding determinants and showed tumor mutations inactivate SMAD4 by disrupting DNA binding or protein stability, linking genotype to molecular defect.

    Evidence Alanine-scanning mutagenesis with in vitro DNA-binding assays; later IHC mutation mapping

    PMID:10871368 PMID:15014009

    Open questions at the time
    • Stability-based inactivation is inferred from protein levels rather than direct turnover measurement
    • MH1 mutation effects assessed in vitro
  9. 2000 High

    Established the timing of DPC4 loss in pancreatic neoplasia, answering when in tumor evolution the suppressor is inactivated.

    Evidence IHC across 188 PanIN lesions correlated with genetic status

    PMID:10766191

    Open questions at the time
    • Correlative; does not prove loss drives progression
    • Restricted to pancreatic lesions
  10. 2003 Medium

    Showed that SMAD4 maintains epithelial identity by suppressing Wnt/β-catenin, explaining transdifferentiation phenotypes upon loss.

    Evidence Mammary-specific conditional knockout with β-catenin IHC and TGF-β1 degradation assays

    PMID:14597578

    Open questions at the time
    • Mechanism linking SMAD4 to β-catenin degradation not biochemically defined
    • Single tissue context
  11. 2004 High

    Identified SCF(β-TrCP1) as a SMAD4 E3 ligase, establishing proteasomal control of SMAD4 abundance and signaling output.

    Evidence Yeast two-hybrid, Co-IP, ubiquitination assays, siRNA, functional readouts

    PMID:14988407

    Open questions at the time
    • Did not define the degron recognized by β-TrCP
    • Upstream priming kinases unidentified at this stage
  12. 2014 High

    Defined the MAPK/GSK3-primed phosphodegron and its antagonism by Wnt, integrating SMAD4 stability into broader signaling crosstalk.

    Evidence Phosphosite mutagenesis, kinase assays, β-TrCP binding, Xenopus gain/loss-of-function

    PMID:25373906

    Open questions at the time
    • In vivo relevance shown in Xenopus; mammalian developmental role not addressed
    • Did not quantify endogenous degradation kinetics
  13. 2014 High

    Demonstrated an essential SMAD4–FOXL2 partnership for FSH synthesis, establishing a tissue-specific transcriptional cofactor relationship.

    Evidence Gonadotrope-specific single and double conditional knockouts with FSH and fertility measurement

    PMID:24739304

    Open questions at the time
    • Direct co-occupancy of the Fshb promoter not shown here
    • Limited to gonadotrope lineage
  14. 2015 Medium

    Showed SMAD4 loss cooperates with PTEN loss to drive metastatic lung tumors via ERRFI1/ErbB2 signaling, expanding its suppressor function to a new epithelium.

    Evidence Pten/Smad4 double knockout mice with transcriptomics, cistromics, pharmacological inhibition

    PMID:25753424

    Open questions at the time
    • Combinatorial context required; single-gene SMAD4 effect not isolated
    • Direct SMAD4 regulation of ERRFI1 not fully resolved
  15. 2017 High

    Established that reversible SMURF2 monoubiquitination, opposed by USP4, tunes SMAD4 activity in stem-cell fate decisions.

    Evidence Ubiquitination assays, Co-IP, USP4 knockdown in ESCs, zebrafish morpholino rescue

    PMID:28468752

    Open questions at the time
    • Monoubiquitination site on SMAD4 not specified here
    • Interplay with degradative ubiquitination not addressed
  16. 2018 High

    Defined an endothelial-specific SMAD4 function preventing arteriovenous malformations via BMP9/ALK1 restraint of PI3K/AKT through CK2, distinguishing a vascular role from canonical transcription.

    Evidence Endothelial-specific inducible knockout with PI3K inhibition, Akt1 deletion, and CK2 manipulation rescues

    PMID:29976569

    Open questions at the time
    • Direct transcriptional repression of CK2 by SMAD4 inferred
    • Human AVM relevance not tested in this study
  17. 2019 High

    Identified ALK-mediated Tyr95 phosphorylation as a DNA-binding-blocking modification, providing a druggable route to restore SMAD4 tumor suppression.

    Evidence In vitro kinase assay, phospho-specific antibody, DNA-binding/reporter assays, ALK inhibition

    PMID:30664791

    Open questions at the time
    • Prevalence of Y95 phosphorylation across tumor types not quantified
    • Structural basis of DNA-binding loss not solved
  18. 2019 Medium

    Showed USP10 deubiquitinates and stabilizes SMAD4 to promote HCC metastasis, illustrating context-dependent oncogenic output of SMAD4 stabilization.

    Evidence siRNA screen, Co-IP, ubiquitination assays, knockdown with SMAD4 rescue, migration assays

    PMID:31721429

    Open questions at the time
    • Single cancer context; reciprocal to suppressor role
    • Ubiquitin linkage type not defined
  19. 2019 High

    Defined a SMAD4–HNF4 feed-forward loop that stabilizes enterocyte identity, explaining how SMAD4 enforces lineage fate.

    Evidence Conditional knockouts, ChIP-seq co-binding, transcriptomics, organoid assays

    PMID:30988513

    Open questions at the time
    • Did not address upstream ligand control of the loop
    • Tumor-suppressive consequence in intestine not directly tested
  20. 2019 Medium

    Identified a SMAD4–FZD4 axis (direct promoter binding plus lncRNA-mediated regulation) coupling SMAD4 to Wnt activation in granulosa cells.

    Evidence ChIP, luciferase, overexpression/knockdown, miRNA sponge assays

    PMID:32415058

    Open questions at the time
    • Single cell-type context
    • lncRNA/miR-29c module not validated in vivo
  21. 2020 Medium

    Showed Wip1 dephosphorylates SMAD4 at Thr277 to restrain its nuclear accumulation and antimitogenic activity, adding phosphatase control to the regulatory layer.

    Evidence Co-IP, phosphatase assay, phospho-specific detection, Xenopus and migration/invasion assays

    PMID:32103600

    Open questions at the time
    • Single lab
    • Relationship of Thr277 to the GSK3/β-TrCP degron not integrated
  22. 2020 Medium

    Revealed a non-transcriptional mitochondrial SMAD4 function binding PKM2 and ATPIF1 to alter metabolism in diabetic nephropathy.

    Evidence Podocyte-specific knockout, subcellular fractionation, Co-IP, enzyme activity and metabolic flux assays

    PMID:31916354

    Open questions at the time
    • Unusual mitochondrial localization not independently confirmed
    • Mechanism of SMAD4 mitochondrial import unknown
  23. 2021 Medium

    Identified FOSL1 de-repression as a sufficient driver of metastasis upon SMAD4 loss in pancreatic cancer.

    Evidence Isogenic SMAD4 cells, in vivo CRISPR/genetic screen, transcriptomics

    PMID:34320363

    Open questions at the time
    • Validation limited to screen confirmation
    • Direct vs indirect FOSL1 repression not resolved
  24. 2021 Medium

    Established SMAD4 as a regulator of adult astrocyte proliferation, extending its role to CNS lineage maintenance.

    Evidence scRNA-seq, conditional deletion, BrdU/clonal analysis, neurosphere assays

    PMID:34549820

    Open questions at the time
    • Downstream transcriptional program in astrocytes unknown
    • Single brain region
  25. 2023 Medium

    Defined PRMT5-mediated R361 methylation as a requirement for SMAD complex assembly and nuclear import, adding methylation to SMAD4 control and linking it to EMT/metastasis.

    Evidence Mass spectrometry, Co-IP, immunofluorescence, R361 mutant functional studies

    PMID:36991117

    Open questions at the time
    • Single lab
    • Interplay with phosphorylation/ubiquitination not addressed
  26. 2023 Medium

    Showed SFPQ sequesters SMAD4 in phase-separated condensates to suppress its activity, introducing biomolecular condensation as a regulatory mechanism.

    Evidence Co-IP, LLPS assays, ChIP, reporter assays, SFPQ phase-separation mutants

    PMID:38103553

    Open questions at the time
    • Physiological/in vivo relevance of condensate sequestration not established
    • Single study

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the layered post-translational marks (phospho-degron, mono-ubiquitination, methylation, tyrosine phosphorylation) and condensate sequestration are coordinated to set SMAD4 output in a given tissue remains unresolved.
  • No integrated model reconciling competing modifications
  • Tissue-specific selectivity of regulators not mapped
  • No structural model of the modified, complex-assembled SMAD4

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0003677 DNA binding 4 GO:0140110 transcription regulator activity 4 GO:0060090 molecular adaptor activity 2
Localization
GO:0005634 nucleus 3 GO:0005739 mitochondrion 1
Pathway
R-HSA-1266738 Developmental Biology 3 R-HSA-162582 Signal Transduction 3 R-HSA-1643685 Disease 3 R-HSA-392499 Metabolism of proteins 3 R-HSA-74160 Gene expression (Transcription) 3
Partners
CBP/P300FAST-1FOXL2HNF4ASFPQSMAD1SMAD2SMAD3
Complex memberships
R-SMAD/SMAD4 heteromeric complexSMAD2/SMAD4/FAST-1 (ARF) complex

Evidence

Reading pass · 32 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1996 DPC4/SMAD4 was identified as a candidate tumor suppressor gene at chromosome 18q21.1, with homozygous deletions in ~30% and intragenic mutations in ~20% of pancreatic carcinomas, implicating it in a TGF-β-like signaling pathway based on sequence similarity to Drosophila Mad. Homozygous deletion mapping, PCR/sequencing of tumor DNA Science High 8553070
1996 DPC4/SMAD4 physically associates with Smad1 in response to BMP and with Smad2 in response to activin or TGF-β, forming regulated heteromeric complexes essential for mesoderm induction and antimitogenic responses in Xenopus embryos and breast epithelial cells. Co-immunoprecipitation, Xenopus embryo overexpression/dominant-negative assays, mammalian cell TGF-β response assays Nature High 8893010
1997 Smad4 is present in the activin-responsive factor (ARF) complex together with FAST-1 and Smad2; Smad4 stabilizes a ligand-stimulated Smad2-FAST-1 complex as an active DNA-binding factor. The FAST-1 C-terminal domain interacts with Smad2 (not Smad4 directly in yeast two-hybrid), but FAST-1 deletion mutants that cannot recruit Smad4 fail to associate with ARF. Co-immunoprecipitation (ligand-regulated), yeast two-hybrid, deletion mutagenesis, DNA-binding assays Nature High 9288972
1997 Smad4 contributes two distinct functions in TGF-β transcriptional complexes: its N-terminal (MH1) domain promotes DNA binding of the Smad2/Smad4/FAST-1 complex, while its C-terminal (MH2) domain provides a transcriptional activation function required for Smad1 or Smad2 to stimulate transcription. Smad4 is not required for nuclear translocation of Smad1/2 or for Smad2-FAST-1 association. Domain deletion and chimera constructs, transcriptional reporter assays, nuclear localization assays in mammalian cells Genes & development High 9389648
1997 Smad3 and Smad4/DPC4 undergo both homomeric and heteromeric interactions via their conserved C-terminal (MH2) domains; Smad4 homomeric interaction additionally requires the N-terminal domain. Cancer-associated mutations in the MH2 domain impair homo- and heteromeric associations and correlate with reduced signaling activity. Yeast two-hybrid, co-immunoprecipitation, transcriptional activation assays in yeast and mammalian cells, analysis of tumor-derived mutations Molecular and cellular biology High 9111321
1998 Smad4/DPC4 and CBP/p300 act as transcriptional coactivators for Smad3 in TGF-β-induced transcriptional activation; CBP associates with the C-terminus of Smad3 in a TGF-β-dependent manner, and this interaction requires Smad4. E1A expression, which blocks CBP function, inhibits TGF-β-induced transcription. Co-immunoprecipitation, transcriptional reporter assays, E1A inhibition experiments Genes & development High 9679060
1997 A 47-amino acid deletion within the middle-linker region of Smad4 abolishes its ability to mediate TGF-β/activin signaling responses, while the N-terminal domain augments ligand-dependent activation, identifying a distinct ligand-response domain in the Smad4 linker. Smad4 deletion/chimera constructs, transcriptional reporter assays in Smad4-null cell line The Journal of biological chemistry Medium 9153220
1998 Homozygous Smad4 knockout mice die before embryonic day 7.5 and fail to gastrulate or form mesoderm. Tetraploid rescue experiments demonstrated that the gastrulation defect is non-cell-autonomous, arising secondary to abnormal visceral endoderm differentiation. Homologous recombination knockout, tetraploid aggregation rescue, histological and molecular marker analysis Genes & development High 9420335
1998 SMAD4 (truncating the C-terminal MH2 domain) homozygous knockout mice fail to undergo endoderm differentiation and mesoderm formation; blastocyst outgrowths show cellular proliferation defects, demonstrating that SMAD4-mediated signals are required for epiblast proliferation, egg-cylinder formation, and mesoderm induction. Homologous recombination knockout (exon 8 truncation), blastocyst outgrowth culture, histological and molecular marker analysis Proceedings of the National Academy of Sciences High 9520423
1997 DPC4/SMAD4 restoration in DPC4-deleted breast tumor cells (MDA-MB-468) reconstitutes TGF-β1-induced growth inhibition and transcriptional activation of a TGF-β sensitive reporter (3TPlux). A DPC4 splice variant lacking residues 223–301 fails to restore TGF-β responsiveness. Transfection/reconstitution in Smad4-null cell line, growth inhibition assays, transcriptional reporter assays Oncogene Medium 9150356
2000 Alanine-scanning mutagenesis of the MH1 domain mapped DNA-binding activity to residues L43–R135, demonstrating that the MH1 domain as a whole is structurally sensitive and that tumor-associated mutations outside the beta-hairpin motif inactivate Smad4 by disrupting DNA binding. Alanine scanning mutagenesis (20 individual mutations), in vitro DNA-binding assays Nucleic acids research High 10871368
2000 Restoration of Smad4 in pancreatic carcinoma cells suppressed tumor formation in vivo without restoring TGF-β sensitivity; instead, Smad4 shifted angiogenic balance by decreasing VEGF expression and increasing thrombospondin-1 expression, reducing vascular density in tumors. Stable reconstitution in Smad4-null pancreatic cancer cells, in vivo nude mouse tumor assays, VEGF/thrombospondin-1 expression analysis Proceedings of the National Academy of Sciences High 10944227
1999 In SW480.7 colon cancer cells, Smad4 loss is due to epigenetic silencing (not mutation); conditional re-expression of Smad4 alone failed to rescue TGF-β antiproliferative responses because co-existing hyperactive Ki-Ras inhibits Smad nuclear accumulation via MAPK phosphorylation. Co-expression of Smad4 with a Ras-phosphorylation-resistant Smad3 (but not wild-type Smad2, Smad3, APC, or TGF-β type II receptor) rescued the antiproliferative response. Ecdysone-inducible Smad4 expression, co-transfection epistasis analysis, cell proliferation assays, p21/c-myc gene response assays The Journal of biological chemistry High 10559252
2004 SCF(β-TrCP1) ubiquitin E3 ligase interacts with Smad4 (but not Smad2, and only indirectly with Smad3 through Smad4) and promotes Smad4 ubiquitination and proteasomal degradation; ectopic SCF(β-TrCP1) inhibited TGF-β-dependent transcriptional activity and cell cycle arrest, while siRNA knockdown of β-TrCP1 increased Smad4 protein levels. Yeast two-hybrid, co-immunoprecipitation, ubiquitination assays, siRNA knockdown, transcriptional reporter assays, cell cycle assays The Journal of biological chemistry High 14988407
2014 Smad4 activity is directly regulated by phosphorylation: FGF activates MAPK which primes three sequential GSK3 phosphorylations in the Smad4 linker region, generating a β-TrCP-bound phosphodegron. Wnt signaling prevents these GSK3 phosphorylations and thereby potentiates TGF-β/Smad4 transcriptional activity. These phosphorylations regulate germ-layer specification in Xenopus embryos. Phosphosite mutagenesis, kinase assays (MAPK, GSK3), Xenopus embryo gain/loss-of-function experiments, β-TrCP binding assays Cell reports High 25373906
2019 ALK tyrosine kinase directly phosphorylates SMAD4 at Tyr95; phospho-Y95 SMAD4 cannot bind DNA and fails to elicit TGF-β gene responses or tumor-suppressive responses. Chemical or genetic inhibition of ALK restores TGF-β responses in ALK-positive tumor cells. In vitro kinase assay, phospho-specific antibody, DNA-binding assay, transcriptional reporter assays, ALK inhibitor treatment and genetic knockdown Nature cell biology High 30664791
2017 USP4 deubiquitinase removes inhibitory monoubiquitination from SMAD4, sustaining its activity in activin/BMP signaling. SMURF2 E3 ligase is recruited to SMAD4 upon ligand-induced R-SMAD-SMAD4 complex formation to add monoubiquitin. The negative regulator c-SKI inhibits SMAD4 monoubiquitination. USP4 depletion in mouse ESCs increased monoubiquitinated SMAD4 and impaired BMP/activin-induced cell fate changes. Ubiquitination assays, co-immunoprecipitation, USP4 knockdown in mouse ESCs, zebrafish morpholino knockdown with rescue experiments The EMBO journal High 28468752
2019 USP10 deubiquitinase directly interacts with Smad4 and stabilizes it by removing proteolytic ubiquitination, thereby activating TGF-β signaling and promoting HCC metastasis. Suppression of USP10 reduced Smad4 protein levels and inhibited HCC cell migration; reconstitution of Smad4 rescued this defect. siRNA library screen, co-immunoprecipitation, ubiquitination assays, shRNA knockdown, Smad4 reconstitution, migration assays Molecular oncology Medium 31721429
2020 Wip1 phosphatase selectively binds and dephosphorylates Smad4 at Thr277 (a key MAPK phosphorylation site), regulating Smad4 nuclear accumulation and protein half-life. Wip1 restrains TGF-β-induced growth arrest, migration, and invasion, and inhibits Smad4 antimitogenic activity in human cells and mesoderm formation in Xenopus. Co-immunoprecipitation, phosphatase assay, phospho-specific antibody detection, Xenopus gain/loss-of-function, cell migration/invasion assays EMBO reports Medium 32103600
2023 PRMT5 interacts with SMAD4 under TGF-β1 treatment and methylates SMAD4 at R361; this methylation is required for SMAD complex formation and nuclear import. SMAD4 R361 mutation abolishes PRMT5-induced EMT and colorectal cancer metastasis. Mass spectrometry, co-immunoprecipitation, immunofluorescence, methylation-specific assays, SMAD4 R361 mutant functional studies Oncogene Medium 36991117
2023 SFPQ, a prion-like RNA-binding protein, physically sequesters Smad4 in liquid-liquid phase separation (LLPS) condensates via its prion-like domain (PrLD), excluding Smad4 from Smad complexes and chromatin, thereby suppressing TGF-β transcriptional responses. SFPQ deficiency or abolition of phase separation activity renders cells hypersensitive to TGF-β. Co-immunoprecipitation, LLPS assays, ChIP, transcriptional reporter assays, SFPQ phase-separation mutants Developmental cell Medium 38103553
2018 Endothelial-specific loss of Smad4 in mice causes arteriovenous malformation (AVM) formation. Mechanistically, BMP9 signaling antagonizes flow-induced AKT activation in an ALK1- and SMAD4-dependent manner; Smad4-deficient endothelial cells display increased PI3K/AKT signaling. BMP9-induced SMAD4 inhibits CK2 (casein kinase 2) transcription, limiting PTEN phosphorylation and AKT activation. PI3K inhibition or endothelial Akt1 deletion rescues AVMs in Smad4-deficient mice. Tamoxifen-inducible endothelial-specific Smad4 knockout mice, pharmacological PI3K inhibition, Akt1 genetic rescue, CK2 inhibition, AKT phosphorylation assays Circulation High 29976569
2019 SMAD4 and HNF4 (HNF4A and HNF4G) function via a reinforcing feed-forward loop in the intestinal epithelium: SMAD4 and HNF4 activate each other's expression and co-bind regulatory elements of differentiation genes to promote and stabilize enterocyte cell identity. Disruption of this HNF4-SMAD4 module results in loss of enterocyte fate in favor of progenitor and secretory cell lineages. Conditional double knockout (HNF4A/G), SMAD4 conditional knockout, ChIP-seq cistromic analysis, transcriptomic profiling, intestinal organoid assays Nature genetics High 30988513
2014 SMAD4 is absolutely required for normal FSH (Fshb) synthesis in vivo; conditional Smad4 deletion in gonadotropes abolishes FSH synthesis. Combined deletion of Smad4 and its DNA-binding cofactor FOXL2 in gonadotropes results in near-complete absence of FSH and female sterility, phenocopying Fshb-knockout mice, establishing SMAD4 and FOXL2 as essential co-regulators of Fshb transcription. Conditional gonadotrope-specific Smad4 and Foxl2 single/double knockout mice, FSH measurement, fertility assessment FASEB journal High 24739304
2020 Under hyperglycemic conditions, Smad4 localizes to mitochondria in podocytes, directly binds the glycolytic enzyme PKM2 (reducing its active tetrameric form) and interacts with ATPIF1 (reducing its degradation), resulting in reduced glycolysis and oxidative phosphorylation and increased ROS production, contributing to diabetic nephropathy. Conditional podocyte-specific Smad4 knockout mice, subcellular fractionation, co-immunoprecipitation, PKM2 activity assays, metabolic flux measurements EMBO reports Medium 31916354
2003 Mammary epithelium-specific Smad4 deletion causes squamous cell carcinoma and mammary abscesses via transdifferentiation. Loss of Smad4 leads to β-catenin accumulation at onset of transdifferentiation; TGF-β1 degrades β-catenin in cultured mammary epithelial cells, but this action is blocked in the absence of Smad4, implicating Smad4 in suppressing Wnt/β-catenin signaling during cell fate maintenance. Cre-loxP conditional mammary-specific Smad4 knockout, immunohistochemistry for β-catenin, TGF-β1 treatment assays in cultured cells Development Medium 14597578
2021 SMAD4 represses FOSL1 expression; in pancreatic cancer cells, SMAD4 loss leads to FOSL1 upregulation that is sufficient to drive metastatic colonization to the lung, identified by an in vivo CRISPR/genetic screen. Isogenic cell lines with/without SMAD4, in vivo functional screen, transcriptomic analysis Cell reports Medium 34320363
2000 DPC4 inactivation in pancreatic cancer occurs late in neoplastic progression: all PanIN-1A, PanIN-1B, and PanIN-2 lesions retained Dpc4 expression, while 31% of PanIN-3 (carcinoma in situ) lesions lost it, demonstrating stage-specific loss of the tumor suppressor. Immunohistochemistry for Dpc4 protein in 188 pancreatic intraepithelial neoplasias (PanINs) correlated with genetic status Cancer research High 10766191
2004 Missense mutations in SMAD4 are concentrated in the MH2 domain (77%), with a mutation cluster region (MCR) at codons 330–370. Mutations outside the MCR correlate with loss of Madh4 protein (suggesting degradation), while MCR mutations retain nuclear protein, indicating that most missense mutations inactivate Smad4 via protein destabilization/degradation rather than direct functional disruption. Sequence analysis of tumors, immunohistochemistry for Madh4 protein in archival cancers with known missense mutations Clinical cancer research Medium 15014009
2015 Combined loss of PTEN and SMAD4 in mouse airway epithelium leads to metastatic adenosquamous lung tumors through ELF3 and ErbB2 pathway activation due to decreased ERRFI1 expression. Combinatorial inhibition of ErbB2 and Akt attenuates tumor progression and invasion. Conditional Pten/Smad4 double knockout mouse model, comparative transcriptomics, in vivo cistromics, pharmacological inhibition Cell reports Medium 25753424
2019 SMAD4 directly binds the FZD4 promoter as a transcription factor to activate FZD4 transcription in granulosa cells, and also promotes FZD4 expression indirectly via induction of a lncRNA (SDNOR) that sponges miR-29c (which would otherwise degrade FZD4 mRNA), establishing a SMAD4-FZD4 axis that activates Wnt signaling to regulate granulosa cell apoptosis. ChIP (SMAD4 binding to FZD4 promoter), luciferase reporter assays, SMAD4 overexpression/knockdown, miRNA sponge assays Cell death & disease Medium 32415058
2021 Smad4 regulates adult astrocyte proliferation in the diencephalon; Smad4 deletion in diencephalic astrocytes reduces in vivo proliferation and in vitro neurosphere formation, identifying Smad4 as a key regulator of adult astrogenesis in this brain region. Single-cell RNA-seq, MACS isolation of astrocytes, conditional Smad4 deletion, BrdU/clonal analysis, neurosphere assays The EMBO journal Medium 34549820

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1996 DPC4, a candidate tumor suppressor gene at human chromosome 18q21.1. Science (New York, N.Y.) 2106 8553070
1996 Partnership between DPC4 and SMAD proteins in TGF-beta signalling pathways. Nature 813 8893010
1998 Mutations in the SMAD4/DPC4 gene in juvenile polyposis. Science (New York, N.Y.) 695 9582123
1996 DPC4 gene in various tumor types. Cancer research 631 8653691
2004 A combined syndrome of juvenile polyposis and hereditary haemorrhagic telangiectasia associated with mutations in MADH4 (SMAD4). Lancet (London, England) 561 15031030
1997 Smad4 and FAST-1 in the assembly of activin-responsive factor. Nature 496 9288972
2018 The role of TGF-β/SMAD4 signaling in cancer. International journal of biological sciences 486 29483830
2000 Loss of expression of Dpc4 in pancreatic intraepithelial neoplasia: evidence that DPC4 inactivation occurs late in neoplastic progression. Cancer research 477 10766191
1998 The tumor suppressor Smad4/DPC4 and transcriptional adaptor CBP/p300 are coactivators for smad3 in TGF-beta-induced transcriptional activation. Genes & development 446 9679060
1998 The tumor suppressor gene Smad4/Dpc4 is required for gastrulation and later for anterior development of the mouse embryo. Genes & development 401 9420335
1997 Dual role of the Smad4/DPC4 tumor suppressor in TGFbeta-inducible transcriptional complexes. Genes & development 393 9389648
2002 Evaluation of candidate genes MAP2K4, MADH4, ACVR1B, and BRCA2 in familial pancreatic cancer: deleterious BRCA2 mutations in 17%. Cancer research 337 12097290
1998 The tumor suppressor SMAD4/DPC4 is essential for epiblast proliferation and mesoderm induction in mice. Proceedings of the National Academy of Sciences of the United States of America 289 9520423
2001 Genetic profile of 22 pancreatic carcinoma cell lines. Analysis of K-ras, p53, p16 and DPC4/Smad4. Virchows Archiv : an international journal of pathology 284 11787853
2000 Immunohistochemical labeling for dpc4 mirrors genetic status in pancreatic adenocarcinomas : a new marker of DPC4 inactivation. The American journal of pathology 271 10623651
2000 Smad4/DPC4-mediated tumor suppression through suppression of angiogenesis. Proceedings of the National Academy of Sciences of the United States of America 223 10944227
1996 Somatic alterations of the DPC4 gene in human colorectal cancers in vivo. Gastroenterology 204 8898652
2004 The prevalence of MADH4 and BMPR1A mutations in juvenile polyposis and absence of BMPR2, BMPR1B, and ACVR1 mutations. Journal of medical genetics 199 15235019
1997 Heteromeric and homomeric interactions correlate with signaling activity and functional cooperativity of Smad3 and Smad4/DPC4. Molecular and cellular biology 189 9111321
2003 Role of Smad4 (DPC4) inactivation in human cancer. Biochemical and biophysical research communications 179 12821112
2002 Aberrant p16(INK4A) and DPC4/Smad4 expression in intraductal papillary mucinous tumours of the pancreas is associated with invasive ductal adenocarcinoma. Gut 150 12010891
1999 Gastric and duodenal polyps in Smad4 (Dpc4) knockout mice. Cancer research 149 10626800
2002 DPC4/Smad4 expression and outcome in pancreatic ductal adenocarcinoma. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 147 12454109
1996 DPC4, a candidate tumor suppressor gene, is altered infrequently in head and neck squamous cell carcinoma. Cancer research 144 8653689
1999 Smad2 and Smad4 gene mutations in hepatocellular carcinoma. Oncogene 137 10490821
1997 DPC4 (SMAD4) mediates transforming growth factor-beta1 (TGF-beta1) induced growth inhibition and transcriptional response in breast tumour cells. Oncogene 136 9150356
1997 Characterization of functional domains within Smad4/DPC4. The Journal of biological chemistry 133 9153220
2002 Juvenile polyposis: massive gastric polyposis is more common in MADH4 mutation carriers than in BMPR1A mutation carriers. Human genetics 128 12136244
1999 Smad4/DPC4 silencing and hyperactive Ras jointly disrupt transforming growth factor-beta antiproliferative responses in colon cancer cells. The Journal of biological chemistry 126 10559252
2019 A reinforcing HNF4-SMAD4 feed-forward module stabilizes enterocyte identity. Nature genetics 125 30988513
2010 Smad4-mediated TGF-beta signaling in tumorigenesis. International journal of biological sciences 122 20087440
2002 Frequent loss of SMAD4/DPC4 protein in colorectal cancers. Gut 117 12077092
2018 SMAD4 Prevents Flow Induced Arteriovenous Malformations by Inhibiting Casein Kinase 2. Circulation 104 29976569
2010 MicroRNA 483-3p suppresses the expression of DPC4/Smad4 in pancreatic cancer. FEBS letters 97 21112326
1999 Mutations of the DPC4/Smad4 gene in neuroendocrine pancreatic tumors. Oncogene 94 10327057
2004 Loss of BMP2, Smad8, and Smad4 expression in prostate cancer progression. The Prostate 92 15042598
2000 Loss of Dpc4 expression in colonic adenocarcinomas correlates with the presence of metastatic disease. The American journal of pathology 92 11021814
2004 Smad4 protein stability is regulated by ubiquitin ligase SCF beta-TrCP1. The Journal of biological chemistry 87 14988407
2011 MicroRNA 421 suppresses DPC4/Smad4 in pancreatic cancer. Biochemical and biophysical research communications 86 21352803
2017 Bmp Induces Osteoblast Differentiation through both Smad4 and mTORC1 Signaling. Molecular and cellular biology 83 27920253
2014 SMAD4 and its role in pancreatic cancer. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 82 25464861
2004 Missense mutations of MADH4: characterization of the mutational hot spot and functional consequences in human tumors. Clinical cancer research : an official journal of the American Association for Cancer Research 81 15014009
2020 SMAD4 and the TGFβ Pathway in Patients with Pancreatic Ductal Adenocarcinoma. International journal of molecular sciences 80 32429474
2015 ErbB2 Pathway Activation upon Smad4 Loss Promotes Lung Tumor Growth and Metastasis. Cell reports 77 25753424
1999 DPC4/SMAD4 mediated tumor suppression of colon carcinoma cells is associated with reduced urokinase expression. Oncogene 76 10340387
2003 Squamous cell carcinoma and mammary abscess formation through squamous metaplasia in Smad4/Dpc4 conditional knockout mice. Development (Cambridge, England) 75 14597578
2019 Deubiquitinating enzyme USP10 promotes hepatocellular carcinoma metastasis through deubiquitinating and stabilizing Smad4 protein. Molecular oncology 74 31721429
2014 The tumor suppressor Smad4/DPC4 is regulated by phosphorylations that integrate FGF, Wnt, and TGF-β signaling. Cell reports 72 25373906
2014 Follicle-stimulating hormone synthesis and fertility depend on SMAD4 and FOXL2. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 70 24739304
1999 Frequent 4-bp deletion in exon 9 of the SMAD4/MADH4 gene in familial juvenile polyposis patients. Genes, chromosomes & cancer 69 10398437
2023 PRMT5 methylating SMAD4 activates TGF-β signaling and promotes colorectal cancer metastasis. Oncogene 68 36991117
2018 Smad4/DPC4. Journal of clinical pathology 66 29720405
2018 SMAD4 Suppresses WNT-Driven Dedifferentiation and Oncogenesis in the Differentiated Gut Epithelium. Cancer research 66 29986996
2024 Significance of TP53, CDKN2A, SMAD4 and KRAS in Pancreatic Cancer. Current issues in molecular biology 63 38666907
2020 SMAD4 activates Wnt signaling pathway to inhibit granulosa cell apoptosis. Cell death & disease 62 32415058
2020 Smad4 promotes diabetic nephropathy by modulating glycolysis and OXPHOS. EMBO reports 60 31916354
1997 Expression and mutational analysis of the DCC, DPC4, and MADR2/JV18-1 genes in neuroblastoma. Cancer research 57 9288786
2000 DCC and SMAD4 alterations in human colorectal and pancreatic tumor dissemination. Oncogene 56 10698524
2000 Cloning and characterization of zebrafish smad2, smad3 and smad4. Gene 53 10767528
2018 Epithelial Smad4 Deletion Up-Regulates Inflammation and Promotes Inflammation-Associated Cancer. Cellular and molecular gastroenterology and hepatology 52 30109253
2019 ALK phosphorylates SMAD4 on tyrosine to disable TGF-β tumour suppressor functions. Nature cell biology 51 30664791
2017 USP4 inhibits SMAD4 monoubiquitination and promotes activin and BMP signaling. The EMBO journal 49 28468752
2017 The DPC4/SMAD4 genetic status determines recurrence patterns and treatment outcomes in resected pancreatic ductal adenocarcinoma: A prospective cohort study. Oncotarget 47 28160547
1999 DPC4/SMAD4 gene alterations in human cancer, and their functional implications. Annals of oncology : official journal of the European Society for Medical Oncology 47 10436786
2022 SMAD4 Suppresses Colitis-associated Carcinoma Through Inhibition of CCL20/CCR6-mediated Inflammation. Gastroenterology 43 35863523
2003 Loss of DPC4 expression and its correlation with clinicopathological parameters in pancreatic carcinoma. World journal of gastroenterology 42 14669329
1997 DPC4 gene mutation in colitis associated neoplasia. Gut 42 9155588
2014 Novel SMAD4 mutation causing Myhre syndrome. American journal of medical genetics. Part A 40 24715504
2000 Homozygous deletions inactivate DCC, but not MADH4/DPC4/SMAD4, in a subset of pancreatic and biliary cancers. Genes, chromosomes & cancer 40 10719364
2021 SMAD4 represses FOSL1 expression and pancreatic cancer metastatic colonization. Cell reports 39 34320363
2002 Smad4 transcriptional pathways and angiogenesis. International journal of gastrointestinal cancer 38 12622415
1998 Smad4 (DPC4)--a potent tumour suppressor? British journal of cancer 37 9862572
2004 Immunohistochemical study of DPC4 and p53 proteins in gallbladder and bile duct cancers. World journal of surgery 36 15573254
2000 No SMAD4 hypermethylation in colorectal cancer. British journal of cancer 35 10993648
2021 Molecular diversity of diencephalic astrocytes reveals adult astrogenesis regulated by Smad4. The EMBO journal 32 34549820
2014 Tak1, Smad4 and Trim33 redundantly mediate TGF-β3 signaling during palate development. Developmental biology 32 25523394
2018 Roles of Smads Family and Alternative Splicing Variants of Smad4 in Different Cancers. Journal of Cancer 31 30410607
2000 Functional mapping of the MH1 DNA-binding domain of DPC4/SMAD4. Nucleic acids research 31 10871368
2000 A novel SMAD4 gene mutation in seminoma germ cell tumors. Cancer research 30 10706106
2023 Smad4 sequestered in SFPQ condensates prevents TGF-β tumor-suppressive signaling. Developmental cell 29 38103553
2019 SMAD4 rare variants in individuals and families with thoracic aortic aneurysms and dissections. European journal of human genetics : EJHG 28 30809044
2000 Gastro-intestinal tumorigenesis in Smad4 mutant mice. Cytokine & growth factor reviews 28 10708962
1999 Higher frequency of DPC4/Smad4 alterations in pancreatic cancer cell lines than in primary pancreatic adenocarcinomas. Cancer letters 27 10408907
1999 Tumor suppressor gene Smad4/DPC4, its downstream target genes, and regulation of cell cycle. Annals of the New York Academy of Sciences 27 10415848
2021 Consequences of Mutations and Abnormal Expression of SMAD4 in Tumors and T Cells. OncoTargets and therapy 26 33888990
2020 Wip1 regulates Smad4 phosphorylation and inhibits TGF-β signaling. EMBO reports 26 32103600
1999 Allelic imbalance in chromosome band 18q21 and SMAD4 mutations in ovarian cancers. Genes, chromosomes & cancer 26 10451707
2019 Smad4 Feedback Enhances BMPR1B Transcription in Ovine Granulosa Cells. International journal of molecular sciences 24 31167348
2011 Keratin 23, a novel DPC4/Smad4 target gene which binds 14-3-3ε. BMC cancer 24 21492476
2001 Nuclear localization of Dpc4 (Madh4, Smad4) in colorectal carcinomas and relation to mismatch repair/transforming growth factor-beta receptor defects. The American journal of pathology 24 11159190
2015 Smad1/5 and Smad4 expression are important for osteoclast differentiation. Journal of cellular biochemistry 23 25711193
2001 Somatic alterations of the DPC4 and Madr2 genes in colorectal cancers and relationship to metastasis. International journal of oncology 23 11172591
2001 Analysis of human meningiomas for aberrations of the MADH2, MADH4, APM-1 and DCC tumor suppressor genes on the long arm of chromosome 18. International journal of cancer 23 11304690
2021 SMAD4 is critical in suppression of BRAF-V600E serrated tumorigenesis. Oncogene 22 34453124
2015 Somatic alterations in juvenile polyps from BMPR1A and SMAD4 mutation carriers. Genes, chromosomes & cancer 22 26171675
2004 Loss of chromosome 18q and DPC4 (Smad4) mutations in appendiceal adenocarcinomas. Oncogene 22 14647445
2009 Smad4: gatekeeper gene in head and neck squamous cell carcinoma. The Journal of clinical investigation 21 19841540
2005 Analysis of SMAD4/DPC4 gene alterations in multiploid colorectal carcinomas. Journal of gastroenterology 21 16082587
2019 SMAD4 Y353C promotes the progression of PDAC. BMC cancer 20 31684910
2008 Expression of DPC4/Smad4 in non-small-cell lung carcinoma and its relationship with angiogenesis. Neoplasma 20 18505344

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