Affinage

ALX4

Homeobox protein aristaless-like 4 · UniProt Q9H161

Length
411 aa
Mass
44.2 kDa
Annotated
2026-06-09
61 papers in source corpus 27 papers cited in narrative 24 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

ALX4 is a paired-like homeodomain transcription factor that controls anterior–posterior patterning and morphogenesis of the developing limb, craniofacial skeleton, skin, and glandular tissues by regulating mesenchymal gene expression (PMID:9374397, PMID:9786416). As a nuclear transcriptional activator, it acts through N-terminal and proline-rich sequences flanking its homeodomain, while the paired-tail is dispensable for activation (PMID:9426253, PMID:9786416). DNA binding is cooperative: ALX4 engages a palindromic TAAT-NNN-ATTA dimer site, with the two molecules making asymmetric protein–protein and protein–DNA contacts, and this dimerization is required for transcriptional activation (PMID:40410151); it also forms DNA-binding heterodimers with the related factor Cart1 that share target sites and developmental roles (PMID:9847249). ALX4 partners with LEF-1 through its N-terminal proline-rich region, co-occupies adjacent sites on the N-CAM promoter, and represses N-CAM, a functional interaction genetically required for embryonic vasculogenesis (PMID:11696550, PMID:16892173). In the limb bud, Alx4 is expressed in anterior mesenchyme and restricts ectopic posterior signaling: its loss causes anterior ectopic Shh, Fgf4, and Hoxd13 expression and preaxial polydactyly, operating in a negative-feedback relationship with Shh and through both Gli3-dependent and Gli3-independent modules (PMID:9374397, PMID:9778501, PMID:16039644, PMID:15968591). ALX4 sits within BMP- and FGF-driven signaling networks—its calvarial expression is induced by BMP via Foxc1 and is partially redundant with Msx2 upstream of Fgfr signaling (PMID:14512019, PMID:15198690)—and it relays FGF-Shp2 signaling by directly binding a conserved Fgf10 intronic enhancer to drive lacrimal gland and eyelid development (PMID:29028795, PMID:25673119). ALX4 additionally suppresses Wnt/β-catenin signaling by promoting GSK3β-dependent β-catenin phosphorylation and degradation (PMID:29183346, PMID:37724761). In humans, ALX4 haploinsufficiency causes parietal foramina, while biallelic loss-of-function and gain-of-function frameshift alleles cause frontonasal dysplasia with skin, hair, and craniofacial defects (PMID:11137991, PMID:11106354, PMID:11017806, PMID:19692347, PMID:37724761).

Mechanistic history

Synthesis pass · year-by-year structured walk · 23 steps
  1. 1997 High

    Established ALX4 as a determinant of limb anterior–posterior identity, answering whether an anterior mesenchymal homeodomain factor restricts posterior signaling-center formation.

    Evidence Targeted knockout mice with molecular marker in situ hybridization (Shh, HoxD13, FGF-4)

    PMID:9374397

    Open questions at the time
    • Direct transcriptional targets in limb not identified
    • Mechanism of ZPA restriction not resolved at molecular level
  2. 1997 Medium

    Confirmed nuclear localization and developmental expression domains, supporting a transcription factor role across craniofacial and limb mesenchyme.

    Evidence Nuclear extract immunoblot, Northern blot, whole-mount in situ hybridization in mouse embryos

    PMID:9426253

    Open questions at the time
    • No target genes identified
    • No DNA-binding specificity defined at this stage
  3. 1997 Medium

    Placed Alx4 within a multigenic anterior digit-patterning network downstream of or parallel to BMP signaling.

    Evidence Bmp4/Alx4 double heterozygous mouse genetic epistasis

    PMID:9268572

    Open questions at the time
    • Biochemical link between Bmp4 and Alx4 not defined
    • Direction of regulation (downstream vs parallel) unresolved
  4. 1998 High

    Defined the negative feedback loop between Alx4 and Shh in limb outgrowth and identified the causative lstJ mutation, distinguishing the feedback from Gli3 control.

    Evidence lstJ allele sequencing, chick Shh/FGF bead implantation, AER removal surgery, in situ hybridization

    PMID:9778501

    Open questions at the time
    • Molecular mediators of Shh suppression of Alx4 unknown
    • Whether feedback is direct or indirect not established
  5. 1998 Medium

    Mapped the transcriptional activation determinants of ALX4 to the N-terminal and proline-rich regions, clarifying domain architecture for activator function.

    Evidence Reporter assays with deletion constructs in cell culture; expression analysis at epithelial–mesenchymal interfaces

    PMID:9786416

    Open questions at the time
    • Coactivators recruited by N-terminal/proline-rich domains not identified
    • Native target promoters not tested
  6. 1999 High

    Demonstrated ALX4–Cart1 heterodimer formation and shared DNA-binding/activation properties, with double mutants revealing functional redundancy in craniofacial and limb development.

    Evidence In vitro DNA binding, gel-shift/Co-IP, reporter assays, double mutant mouse analysis

    PMID:9847249

    Open questions at the time
    • In vivo distribution of homo- vs heterodimers unknown
    • Distinct vs shared target genes not enumerated
  7. 2001 High

    Identified LEF-1 as a direct ALX4 partner and N-CAM as a regulated target, linking ALX4 to promoter co-occupancy and gene repression.

    Evidence Co-IP with domain mapping, N-CAM promoter reporter, endogenous N-CAM immunoblot in primary stromal cells

    PMID:11696550

    Open questions at the time
    • Whether the interaction modulates Wnt signaling at this stage not addressed
    • Genome-wide co-targets unknown
  8. 2001 High

    Showed Alx3/Alx4 cooperate to suppress apoptosis in frontonasal mesenchyme, establishing redundancy in craniofacial outgrowth.

    Evidence Alx3 null and Alx3/Alx4 double mutant mice, histology, TUNEL assay

    PMID:11641221

    Open questions at the time
    • Pro-survival target genes not identified
    • Mechanism linking ALX factors to apoptosis suppression unknown
  9. 2001 High

    Established ALX4 as a dosage-sensitive human gene, with haploinsufficiency causing parietal foramina.

    Evidence Human mutation analysis, loss-of-function identification, FISH mapping across multiple families

    PMID:11017806 PMID:11106354 PMID:11137991

    Open questions at the time
    • Calvarial target genes not identified
    • Threshold for dosage sensitivity not defined
  10. 2003 Medium

    Positioned Alx4 downstream of BMP and Foxc1 in calvarial osteoprogenitor induction.

    Evidence Foxc1 null mice, BMP bead implantation, in situ hybridization, BrdU assay

    PMID:14512019

    Open questions at the time
    • Whether Foxc1 directly regulates the Alx4 promoter not shown
    • Direct BMP-responsive elements not mapped
  11. 2004 Medium

    Defined partial redundancy and mutual regulation between Alx4 and Msx2, placing them upstream of Fgfr signaling in skull ossification.

    Evidence Alx4/Msx2 compound mutant mice, multi-marker in situ hybridization, alkaline phosphatase staining

    PMID:15198690

    Open questions at the time
    • Direct vs indirect mutual regulation unresolved
    • Whether Fgfr1/2 are direct targets not established
  12. 2005 High

    Dissected Gli3-dependent and Gli3-independent control of Alx4, showing early ectopic Fgf4/Hoxd13 is SHH-independent while polydactyly requires Alx4.

    Evidence Alx4/Gli3 double mutants, reporter analysis in Shh-/- and Gli3-/- backgrounds, in situ hybridization

    PMID:15968591 PMID:16039644

    Open questions at the time
    • Enhancer elements mediating Gli3-dependent control not mapped
    • Mechanism of SHH-independent ectopic gene expression unknown
  13. 2006 High

    Established a stromal-cell-autonomous role for Alx4 in mammary ductal morphogenesis, induced by estradiol and linked to MMP balance.

    Evidence Alx4 null analysis, fat pad transplantation/cell mixing, RT-PCR for MMPs/HGF

    PMID:16916507

    Open questions at the time
    • Whether MMP2/MMP9 are direct transcriptional targets unknown
    • Estradiol-to-Alx4 induction mechanism not defined
  14. 2006 Medium

    Genetically validated the Alx4–LEF-1 interaction in vivo, showing it is required for embryonic vasculogenesis.

    Evidence Lef1-/-/Alx4lstD/lstD double mutant embryos, PECAM staining

    PMID:16892173

    Open questions at the time
    • Vascular target genes of the Alx4–LEF-1 complex not identified
    • Cell type driving the vascular defect not pinpointed
  15. 2009 Medium

    Linked biallelic ALX4 loss to a broader human phenotype including skin and hair follicle defects, defining tissue-level requirements.

    Evidence Homozygosity mapping, p.R265X mutation identification, NMD testing, skin biopsy differentiation analysis

    PMID:19692347

    Open questions at the time
    • Epidermal target genes not identified
    • Single family limits genotype–phenotype generalization
  16. 2013 Medium

    Extended Alx4 function to genital tubercle development, linking it to Fibronectin-dependent cell migration and Hh/Gli3 interaction.

    Evidence Alx4Lst/Lst and combinatorial mutants, cell migration labeling, Fibronectin immunostaining, Hh-pathway in situ

    PMID:23942202

    Open questions at the time
    • Whether Fibronectin is a direct target unknown
    • Mechanism of migration control not defined
  17. 2015 Medium

    Identified Fgf10 as the effector through which Alx4 controls eyelid fusion, connecting it to periderm phospho-c-Jun signaling.

    Evidence Spontaneous Alx4 allele, Fgf10 in situ hybridization, phospho-c-Jun immunostaining

    PMID:25673119

    Open questions at the time
    • Direct binding to Fgf10 regulatory elements not shown in this study
    • Link to phospho-c-Jun mechanistically indirect
  18. 2015 Medium

    Revealed an oncogenic context where ALX4 complexes with HOXB13 to drive EMT and invasion via SLUG.

    Evidence Co-IP, overexpression/siRNA, invasion assays, SLUG knockdown rescue in ovarian cancer cells

    PMID:25944620

    Open questions at the time
    • Whether SLUG is a direct ALX4/HOXB13 target not shown
    • Reciprocal Co-IP validation not described
  19. 2017 High

    Demonstrated direct ALX4 binding to a conserved Fgf10 intronic enhancer downstream of FGF-Shp2 signaling, defining ALX4 as a signaling relay for lacrimal gland development.

    Evidence ChIP/reporter assay, neural-crest Shp2 conditional knockout, in situ hybridization, human genetic analysis

    PMID:29028795

    Open questions at the time
    • Other direct enhancer targets not catalogued
    • Cofactors at the Fgf10 enhancer not identified
  20. 2017 Medium

    Defined ALX4 as a suppressor of Wnt/β-catenin signaling via GSK3β-dependent β-catenin degradation, with tumor-suppressive consequences.

    Evidence Wnt reporter assay, phospho-β-catenin western blot, overexpression, nude mouse xenograft

    PMID:29183346

    Open questions at the time
    • Mechanism by which a transcription factor promotes GSK3β-dependent degradation not resolved
    • Direct vs indirect effect on the destruction complex unknown
  21. 2023 Medium

    Identified a gain-of-function tail-elongating frameshift causing dominant frontonasal dysplasia and confirmed ALX4 negatively regulates Wnt/β-catenin in patient cells.

    Evidence Whole-exome sequencing, transcriptional reporter assay, Wnt target gene expression in patient keratinocytes

    PMID:37724761

    Open questions at the time
    • Mechanism of tail-domain-driven hyperactivation not defined
    • Single family limits generalization
  22. 2024 High

    Resolved the structural basis of cooperative ALX4 dimer binding to the TAAT-NNN-ATTA palindrome and showed dimerization is required for activation, stratifying disease variants by molecular defect.

    Evidence Protein structure determination, mutant in vitro DNA binding, reporter assays in cranial neural crest cells, mutagenesis

    PMID:40410151

    Open questions at the time
    • In vivo genome-wide binding map not established
    • Functional consequences of each disease variant not tested in animals
  23. 2024 Medium

    Used lineage-specific inactivation to assign ALX4 craniofacial and hair phenotypes primarily to cranial neural crest, revealing partly redundant roles across lineages.

    Evidence Conditional Alx4 knockout with neural crest and cranial mesoderm Cre lines, phenotypic analysis

    PMID:38481039

    Open questions at the time
    • Lineage-specific target genes not identified
    • Redundant factors in each lineage not defined

Open questions

Synthesis pass · forward-looking unresolved questions
  • The genome-wide direct target repertoire of ALX4 and the molecular basis by which it links DNA-binding to Wnt destruction-complex activity and FGF/SHH feedback remain unresolved.
  • No comprehensive in vivo ChIP-seq target map
  • Mechanism connecting ALX4 to GSK3β-dependent β-catenin degradation undefined
  • How dimer cooperativity is tuned by partners (LEF-1, Cart1, HOXB13) across tissues unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140110 transcription regulator activity 4 GO:0003677 DNA binding 3
Localization
GO:0005634 nucleus 1
Pathway
R-HSA-1266738 Developmental Biology 4 R-HSA-162582 Signal Transduction 4 R-HSA-1643685 Disease 3 R-HSA-74160 Gene expression (Transcription) 3
Partners
CART1HOXB13LEF1

Evidence

Reading pass · 24 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1997 ALX4 (Alx-4) encodes a paired-type homeodomain protein expressed in anterior limb bud mesenchyme; homozygous null mice develop preaxial polydactyly associated with ectopic anterior ZPA formation, as shown by anterior expression of Sonic hedgehog, HoxD13, and FGF-4, establishing ALX4 as a determinant of anterior-posterior positional identity that restricts ZPA formation to the posterior limb bud mesenchyme. Targeted gene disruption (knockout mice), whole-mount in situ hybridization for Shh, HoxD13, FGF-4, HoxB8, Gli3; chromosomal mapping Development (Cambridge, England) High 9374397
1997 Alx4 protein is found in nuclear extracts of mouse embryos, consistent with its function as a transcription factor; Northern blot and whole-mount in situ hybridization localize Alx4 expression to craniofacial mesenchyme, first branchial arch, and limb bud during development. Northern blot, whole-mount in situ hybridization, immunoblot of nuclear extracts with anti-Alx4 antibodies Gene Medium 9426253
1997 Genetic interaction between Bmp4, Gli3, and Alx4 in anterior digit patterning: double heterozygotes of Bmp4 and Alx4 null alleles display ectopic anterior digits specifically on hindlimbs, placing Alx4 downstream or in parallel with Bmp4 signaling in a multigenic control network for anterior digit formation. Genetic epistasis — double heterozygous mutant mice (Bmp4+/-; Alx4+/-), phenotypic analysis Developmental biology Medium 9268572
1998 A 16 bp deletion in the homeobox region of Alx-4 in Strong's Luxoid (lstJ) mice causes a frameshift and protein truncation, identifying this as the causative mutation for polydactyly in lstJ mice; chick Alx-4 expression is complementary to Shh, and Shh/FGF application suppresses Alx-4 expression while AER removal experiments indicate a negative feedback loop between Alx-4 and Shh during limb outgrowth, independent of Gli3. Sequence analysis of lstJ allele, chick Alx-4 cloning, bead implantation (Shh/FGF protein application), AER removal surgery, in situ hybridization in polydactylous mutants Development (Cambridge, England) High 9778501
1998 Alx-4 is a potent transcriptional activator when expressed in cell culture; optimal transcriptional activation requires specific sequences in the N-terminal region and a proline-rich domain downstream of the paired-like homeodomain, but not the paired-tail (C terminus). Alx-4 is expressed in mesenchymal condensations at sites of epithelial-mesenchymal interaction (osteoblast precursors, dermal papilla of hair/whisker follicles, dental papilla, mammary gland mesenchyme). Reporter gene (transcriptional activation) assays in cell culture with deletion constructs; whole-mount in situ hybridization; immunofluorescence Developmental dynamics : an official publication of the American Association of Anatomists Medium 9786416
1999 ALX4 and Cart1 (paired-type homeodomain proteins) form DNA-binding heterodimers in vitro with similar binding activity to palindromic elements; they similarly activate transcription from reporter genes containing high-affinity binding sites in cell culture; double mutant mice show additive/exacerbated craniofacial and limb defects (polydactyly, nasal cartilage fusion failure, split sternum), demonstrating functional redundancy and both unique and shared developmental roles. In vitro DNA binding assay, co-immunoprecipitation/gel-shift (heterodimer formation), reporter gene assay in cell culture, double mutant mouse genetic analysis Development (Cambridge, England) High 9847249
2001 Alx4 physically interacts with LEF-1 through a proline-rich domain in the N-terminal region of Alx4 and the HMG-box DNA-binding domain of LEF-1; ALX4 and LEF-1 can bind simultaneously to adjacent sites on the N-CAM promoter, altering N-CAM promoter activity; expression of Alx4 in primary mammary stromal cells decreases endogenous N-CAM protein levels. Co-immunoprecipitation, domain deletion mutants, reporter gene assay on N-CAM promoter, immunoblot of endogenous N-CAM in primary cells The Journal of biological chemistry High 11696550
2001 Alx3/Alx4 double mutant mice develop severe nasal clefting and craniofacial defects absent in single mutants; increased apoptosis localized to the frontonasal process in E10.0 double mutants establishes that Alx3 and Alx4 cooperate to suppress apoptosis in outgrowing frontonasal mesenchyme, with functional redundancy in craniofacial development. Generation of Alx3 null (lacZ knock-in) and double mutant mice, histological and anatomical analysis, TUNEL apoptosis assay Development (Cambridge, England) High 11641221
2001 Haploinsufficiency of ALX4 (a paired-related homeodomain transcription factor at 11p11-p12) causes parietal foramina (skull ossification defects) in humans, establishing ALX4 as a dosage-sensitive regulator of calvarial bone formation. Mutation analysis in human patients with parietal foramina; identification of loss-of-function mutations in ALX4; FISH mapping of 11p11.2 deletions Nature genetics High 11017806 11106354 11137991
2003 Foxc1 regulates BMP-mediated induction of Alx4 (and Msx2) in calvarial mesenchyme; BMP induces Alx4 expression in this tissue, and this induction requires the forkhead transcription factor Foxc1; loss of Foxc1 reduces Alx4 and Msx2 expression and impairs osteoprogenitor cell proliferation. Analysis of ch (Foxc1 null) mutant mice, BMP bead implantation, in situ hybridization, BrdU proliferation assay Developmental biology Medium 14512019
2004 Alx4 and Msx2 are partially functionally redundant in skull vault ossification; incremental loss of alleles causes additive exacerbation of skull defects; in Msx2 null mice, Alx4 expression is decreased (but not abolished) in the coronal suture region, and vice versa, indicating mutual regulation; expression of Fgfr1 and Fgfr2 (but not Twist1 or Runx2) is reduced in both single mutants, placing them upstream of Fgfr signaling in the osteogenic network. Alx4/Msx2 double mutant mouse generation and compound genotype analysis, in situ hybridization for multiple markers, alkaline phosphatase staining Journal of anatomy Medium 15198690
2005 In Alx4 null limb buds, anterior ectopic expression of Fgf4 and Hoxd13 occurs independently of SHH signaling at early stages, while later polydactyly requires SHH; Gli3-dependent and Gli3-independent modules control Alx4 expression in the limb bud (total absence of reporter in Gli3-/- background; expansion in Shh-/- background), and loss of the severe polydactyly in Gli3-/-;Alx4-/- double mutants shows that this polydactyly requires Alx4 function. Alx4/Gli3 double mutant analysis, reporter construct expression analysis in Shh-/- and Gli3-/- backgrounds, in situ hybridization Developmental biology High 15968591 16039644
2006 Alx4 is expressed in mammary stromal cells adjacent to terminal end buds during puberty; its expression is induced by 17β-estradiol in stromal cells; loss of Alx4 causes defective ductal morphogenesis (delayed development, distorted duct size, reduced branching). The morphogenic defect is stromal-cell-autonomous: Alx4-deficient stromal cells combined with wild-type epithelial cells recapitulate the defect, but wild-type stromal cells rescue Alx4-deficient epithelial cells. MMP2 is increased 40% and MMP9 decreased 50% in Alx4-deficient mammary stromal cells. Alx4 null mouse analysis, mammary fat pad transplantation (stromal/epithelial mixing experiments), whole-mount analysis, RT-PCR for HGF/MMP2/MMP3/MMP9 Developmental biology High 16916507
2006 Genetic interaction between Lef1 and Alx4 is required for early embryonic development; compound Lef1-/-/Alx4lstD/lstD double mutant mice die by E9.5 (whereas single mutants survive), with defective vasculogenesis in embryonic and extraembryonic tissues, genetically confirming the in vitro Alx4–LEF-1 interaction. Double mutant mouse generation (Lef1-/-/Alx4lstD/lstD), embryo analysis, PECAM staining for vasculature The International journal of developmental biology Medium 16892173
2009 A homozygous nonsense mutation in ALX4 (p.R265X) truncating the homeodomain and paired-tail domain produces a non-functional protein (mRNA is stable; no NMD); in patient skin, there is a hypomorphic interfollicular epidermis with reduced suprabasal layers, impaired interfollicular epidermal differentiation, and altered hair follicle differentiation, establishing ALX4 as required for craniofacial, skin, and hair follicle development. Homozygosity mapping, mutation identification, RT-PCR (NMD testing), skin biopsy histology and differentiation marker analysis Human molecular genetics Medium 19692347
2014 NMR solution structures of the ALX4 homeodomain were determined, providing structural coverage of the DNA-binding domain. Solution NMR structure determination Journal of structural and functional genomics Medium 24941917
2015 HOXB13 and ALX4 form a protein complex in ovarian cancer cells; exogenous expression of either promotes EMT and invasion, while depletion suppresses invasion and reverses EMT; both HOXB13 and ALX4 promote SLUG expression, and SLUG is required for their pro-EMT and pro-invasion effects. Co-immunoprecipitation (complex formation), overexpression and siRNA knockdown, invasion assays, SLUG knockdown rescue experiments, immunoblot Oncotarget Medium 25944620
2017 ALX4 suppresses the Wnt/β-catenin pathway by promoting GSK3β-dependent phosphorylation and degradation of β-catenin in breast cancer cells; ectopic ALX4 expression inhibits cell proliferation and metastasis in vitro and in vivo. Luciferase reporter assay (Wnt/β-catenin), Western blot for phospho-β-catenin/total β-catenin, overexpression in cancer cell lines, nude mouse xenograft Journal of experimental & clinical cancer research : CR Medium 29183346
2017 Alx4 directly controls Fgf10 expression in periocular mesenchyme by binding a conserved intronic enhancer element of Fgf10 (conserved in terrestrial but not aquatic animals); Alx4 expression in the neural crest requires Shp2-mediated FGF signaling; loss of Alx4/ALX4 causes lacrimal gland aplasia in mouse and human, positioning Alx4 as a relay in an FGF-Shp2-FGF signaling cascade. ChIP/reporter assay (Alx4 binding to Fgf10 intronic element), conditional knockout mice (Shp2 in neural crest), in situ hybridization, human patient genetic analysis PLoS genetics High 29028795
2015 Loss of Alx4 causes reduced Fgf10 expression in eyelid mesenchyme and failure of eyelid fusion in mice; reduced Fgf10 is accompanied by a decreased number of periderm cells expressing phospho-c-Jun, establishing that Alx4 regulates eyelid fusion through control of Fgf10 expression. Novel Alx4 spontaneous allele characterization, in situ hybridization for Fgf10, immunostaining for phospho-c-Jun in periderm Mammalian genome : official journal of the International Mammalian Genome Society Medium 25673119
2023 A frameshift insertion in ALX4 (c.985_986insGTGC, p.Pro329Argfs*115) that elongates the protein tail domain causes dominant frontonasal dysplasia with ectodermal defects; using a reporter assay, the elongated ALX4 protein shows increased transcriptional activity (gain-of-function); patient keratinocytes show altered expression of Wnt/β-catenin pathway genes, consistent with ALX4 negatively regulating this pathway. Whole-exome sequencing, reporter assay for ALX4 transcriptional activity, Wnt/β-catenin target gene expression in patient keratinocytes American journal of medical genetics. Part A Medium 37724761
2024 Crystal/structural analysis of the ALX4 dimer reveals that ALX4 binds a TAAT-NNN-ATTA palindromic dimer site; seven residues participate in dimer binding (conserved across Paired-like family but not other homeodomain proteins); the two ALX4 molecules within the dimer use distinct residues to form asymmetric protein-protein and protein-DNA contacts; ALX4 cooperativity (dimerization) is required for transcriptional activation; disease variants cause distinct molecular defects including loss of cooperativity; ALX4 binds this motif independently of TWIST1 in cranial neural crest cells. Protein structure determination (crystal/cryo), in vitro DNA binding assays with mutant ALX4 variants, reporter gene assay in cranial neural crest cells, active-site mutagenesis Nature communications High 40410151
2024 Tissue-specific Cre-mediated inactivation of Alx4 in cranial neural crest recapitulates craniofacial defects of Alx4-null mice; Alx4 inactivation in cranial neural crest causes restricted hair loss over anterior skull, while inactivation in cranial mesoderm does not affect hair, revealing that Alx4 plays partly redundant roles in multiple lineages during hair follicle development. Conditional knockout mice (Alx4f/f with lineage-specific Cre lines for neural crest and cranial mesoderm), phenotypic analysis Developmental dynamics : an official publication of the American Association of Anatomists Medium 38481039
2013 Alx4 is required for normal genital tubercle (GT) development; Alx4 loss-of-function (Alx4Lst/Lst) results in hypoplasia of the dorsal GT and reduced Fibronectin expression; cell migration from infra-umbilical mesenchyme toward the dorsal GT is impaired; augmented Hh signaling-related gene expression in Alx4 mutants, and combinatorial mutant analysis places Alx4 in a genetic interaction with Shh and Gli3 during GT formation. Alx4Lst/Lst mutant and combinatorial mutant analysis (with Shh, Gli3), tissue labeling for cell migration, in situ hybridization for Hh pathway genes, immunostaining for Fibronectin European journal of human genetics : EJHG Medium 23942202

Source papers

Stage 0 corpus · 61 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1997 Haploinsufficient phenotypes in Bmp4 heterozygous null mice and modification by mutations in Gli3 and Alx4. Developmental biology 243 9268572
1997 Polydactyly and ectopic ZPA formation in Alx-4 mutant mice. Development (Cambridge, England) 170 9374397
2010 Performance of epigenetic markers SEPT9 and ALX4 in plasma for detection of colorectal precancerous lesions. PloS one 147 20140221
2001 Severe nasal clefting and abnormal embryonic apoptosis in Alx3/Alx4 double mutant mice. Development (Cambridge, England) 147 11641221
2003 Alx1, a member of the Cart1/Alx3/Alx4 subfamily of Paired-class homeodomain proteins, is an essential component of the gene network controlling skeletogenic fate specification in the sea urchin embryo. Development (Cambridge, England) 146 12756175
2001 Haploinsufficiency of the human homeobox gene ALX4 causes skull ossification defects. Nature genetics 114 11137991
1999 Physical and genetic interactions between Alx4 and Cart1. Development (Cambridge, England) 113 9847249
2003 Progression of calvarial bone development requires Foxc1 regulation of Msx2 and Alx4. Developmental biology 105 14512019
2009 ALX4 dysfunction disrupts craniofacial and epidermal development. Human molecular genetics 100 19692347
2000 The ALX4 homeobox gene is mutated in patients with ossification defects of the skull (foramina parietalia permagna, OMIM 168500). Journal of medical genetics 96 11106354
1998 The role of Alx-4 in the establishment of anteroposterior polarity during vertebrate limb development. Development (Cambridge, England) 88 9778501
2000 Haploinsufficiency of ALX4 as a potential cause of parietal foramina in the 11p11.2 contiguous gene-deletion syndrome. American journal of human genetics 70 11017806
2006 Enlarged parietal foramina caused by mutations in the homeobox genes ALX4 and MSX2: from genotype to phenotype. European journal of human genetics : EJHG 57 16319823
1998 Alx-4, a transcriptional activator whose expression is restricted to sites of epithelial-mesenchymal interactions. Developmental dynamics : an official publication of the American Association of Anatomists 52 9786416
2005 Function and regulation of Alx4 in limb development: complex genetic interactions with Gli3 and Shh. Developmental biology 47 16039644
2001 Alx4 binding to LEF-1 regulates N-CAM promoter activity. The Journal of biological chemistry 45 11696550
1997 Alx-4: cDNA cloning and characterization of a novel paired-type homeodomain protein. Gene 45 9426253
2004 Alx4 and Msx2 play phenotypically similar and additive roles in skull vault differentiation. Journal of anatomy 37 15198690
2015 HOXB13 and ALX4 induce SLUG expression for the promotion of EMT and cell invasion in ovarian cancer cells. Oncotarget 36 25944620
2017 ALX4, an epigenetically down regulated tumor suppressor, inhibits breast cancer progression by interfering Wnt/β-catenin pathway. Journal of experimental & clinical cancer research : CR 34 29183346
2005 Genetic interaction of Gli3 and Alx4 during limb development. The International journal of developmental biology 28 15968591
2004 Malformation of cortical and vascular development in one family with parietal foramina determined by an ALX4 homeobox gene mutation. AJNR. American journal of neuroradiology 26 15569759
2019 Epigenetic silencing of ALX4 regulates microcystin-LR induced hepatocellular carcinoma through the P53 pathway. The Science of the total environment 25 31132711
2011 Mild nasal malformations and parietal foramina caused by homozygous ALX4 mutations. American journal of medical genetics. Part A 25 22140057
2017 Alx4 relays sequential FGF signaling to induce lacrimal gland morphogenesis. PLoS genetics 23 29028795
2013 Vertical transmission of a frontonasal phenotype caused by a novel ALX4 mutation. American journal of medical genetics. Part A 22 23401352
2013 Genetic analysis of the role of Alx4 in the coordination of lower body and external genitalia formation. European journal of human genetics : EJHG 21 23942202
2001 Familial case of Potocki-Shaffer syndrome associated with microdeletion of EXT2 and ALX4. Clinical genetics 21 11903336
2015 Methylation pattern of ALX4 gene promoter as a potential biomarker for blood-based early detection of colorectal cancer. Advanced biomedical research 20 26918234
2019 Downregulated long noncoding RNA LINC00313 inhibits the epithelial-mesenchymal transition, invasion, and migration of thyroid cancer cells through inhibiting the methylation of ALX4. Journal of cellular physiology 19 31093972
2006 Loss of Alx4, a stromally-restricted homeodomain protein, impairs mammary epithelial morphogenesis. Developmental biology 16 16916507
2020 Overexpression of circ_0001445 decelerates hepatocellular carcinoma progression by regulating miR-942-5p/ALX4 axis. Biotechnology letters 13 32856218
2013 Potocki-Shaffer deletion encompassing ALX4 in a patient with frontonasal dysplasia phenotype. American journal of medical genetics. Part A 12 24376213
2010 HoxB2, HoxB4 and Alx4 genes are downregulated in the cadmium-induced omphalocele in the chick model. Pediatric surgery international 12 20625746
2019 miR-1470 regulates cell proliferation and apoptosis by targeting ALX4 in hepatocellular carcinoma. Biochemical and biophysical research communications 11 31791584
2018 Association of Novel ALX4 Gene Polymorphisms with Antidepressant Treatment Response: Findings from the CO-MED Trial. Molecular neuropsychiatry 11 29998114
2015 A 20 bp Duplication in Exon 2 of the Aristaless-Like Homeobox 4 Gene (ALX4) Is the Candidate Causative Mutation for Tibial Hemimelia Syndrome in Galloway Cattle. PloS one 11 26076463
2014 Mild nasal clefting may be predictive for ALX4 heterozygotes. American journal of medical genetics. Part A 11 24764194
2009 Loss of ALX4 expression in epithelial cells and adjacent stromal cells in breast cancer. Journal of clinical pathology 10 19783719
2006 Genetic interaction between Lef1 and Alx4 is required for early embryonic development. The International journal of developmental biology 10 16892173
2021 circKLHL24 Blocks Breast Cancer Development by Regulating the miR-1204/ALX4 Network. Cancer biotherapy & radiopharmaceuticals 9 33781094
2015 A novel allele of Alx4 results in reduced Fgf10 expression and failure of eyelid fusion in mice. Mammalian genome : official journal of the International Mammalian Genome Society 9 25673119
2010 [Hypermethylation of the CDH1, SEPT9, HLTF and ALX4 genes and their diagnostic significance in colorectal cancer]. Voprosy onkologii 9 20552891
2021 Circular RNA Circ_0000098 Elevates ALX4 Expression via Adsorbing miR-1204 to Inhibit the Progression of Hepatocellular Carcinoma. Frontiers in oncology 7 34900665
2018 Identification of a novel homozygous ALX4 mutation in two unrelated patients with frontonasal dysplasia type-2. American journal of medical genetics. Part A 7 29681084
2020 A Novel Missense Variant in the ALX4 Gene Underlies Mild to Severe Frontonasal Dysplasia in a Consanguineous Family. Genetic testing and molecular biomarkers 5 32216639
2020 Downregulation of microRNA-324-3p inhibits lung cancer by blocking the NCAM1-MAPK axis through ALX4. Cancer gene therapy 5 33087824
2013 Multiple abnormalities due to a nonsense mutation in the Alx4 gene. Genetics and molecular research : GMR 4 23979902
2021 A transgenic Alx4-CreER mouse to analyze anterior limb and nephric duct development. Developmental dynamics : an official publication of the American Association of Anatomists 3 33728725
2006 The bovine aristaless-like homeobox 4 (ALX4) as a candidate gene for syndactyly. Cytogenetic and genome research 3 17065792
2025 The ALX4 dimer structure provides insight into how disease alleles impact function. Nature communications 2 40410151
2024 Lineage-specific requirements of Alx4 function in craniofacial and hair development. Developmental dynamics : an official publication of the American Association of Anatomists 2 38481039
2021 De novo ALX4 variant detected in child with non-syndromic craniosynostosis. Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas 2 34586326
2020 Vertical transmission of a large calvarial ossification defect due to heterozygous variants of ALX4 and TWIST1. American journal of medical genetics. Part A 1 33369125
2014 Solution NMR structures of homeodomains from human proteins ALX4, ZHX1, and CASP8AP2 contribute to the structural coverage of the Human Cancer Protein Interaction Network. Journal of structural and functional genomics 1 24941917
2026 Mir-301b-3p Targets Alx4 to Suppress Cisplatin Sensitivity in Breast Cancer through DNA Damage. Advanced biology 0 41944225
2025 piR-19521 facilitates cancer stem cell-like properties by enhancing ALX4 transcription via an enhancer RNA-like mechanism in colorectal cancer. International journal of biological macromolecules 0 41109384
2024 A rare homozygous ALX4 mutation in a Bangladeshi girl with frontonasal dysplasia type-2 (FND2). Heliyon 0 39157323
2024 Correction: A 20 bp Duplication in Exon 2 of the Aristaless-Like Homeobox 4 Gene (ALX4) Is the Candidate Causative Mutation for Tibial Hemimelia Syndrome in Galloway Cattle. PloS one 0 39298916
2023 Dominant frontonasal dysplasia with ectodermal defects results from increased activity of ALX4. American journal of medical genetics. Part A 0 37724761
2016 [Mother and son with enlarged parietal foramina, persistent fetal vein, and ALX4 mutation]. No to hattatsu = Brain and development 0 27349084

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