| 1996 |
LDB1 (Ldb1/NLI) was identified as a LIM-domain-binding factor that interacts with high affinity with the LIM domains of LIM-homeodomain proteins (e.g., Lhx1/Lim1). High-affinity binding requires paired LIM domains and is restricted to LIM-HD and LMO subgroup LIM domains. In Xenopus embryos, XLdb1 synergizes with Xlim-1 to form partial secondary axes and activate downstream genes, demonstrating a physical and functional interaction. |
Yeast two-hybrid isolation, in vitro binding assays, Xenopus embryo microinjection with gain-of-function rescue |
Nature |
High |
8918878
|
| 1997 |
In erythroid cells, LDB1/NLI assembles into a multiprotein DNA-binding complex with LMO2, TAL1, E2A/E47, and GATA-1. This oligomeric complex binds a bipartite DNA motif comprising an E-box (CAGGTG) followed ~9 bp downstream by a GATA site, and functions as a transcriptional transactivating complex. All five proteins are required for in vivo assembly. |
Co-immunoprecipitation, electrophoretic mobility shift assay (EMSA), in vivo assembly assays in erythroid cells |
The EMBO journal |
High |
9214632
|
| 1997 |
A 38-amino-acid C-terminal fragment of NLI/LDB1 is sufficient for association with nuclear LIM domains. NLI forms high-affinity homodimers through its N-terminal 200 amino acids; dimerization is not required for LIM domain association. NLI formed complexes with Lmx1 on the rat insulin I promoter and inhibited LIM domain-dependent synergistic transcriptional activation, indicating NLI can act as a negative regulator of synergistic transcriptional responses. |
Deletion mutagenesis, in vitro binding assays, chemical cross-linking, reporter gene assays in transfected cells |
Molecular and cellular biology |
High |
9315627
|
| 1997 |
Ldb1 and LMO2 form a stable complex in murine erythroleukemia cells, with binding affinity greater than between LMO2 and SCL. The C-terminal 76 residues of Ldb1 are sufficient for LMO2 interaction. Ldb1, LMO2, and SCL/E12 can assemble as a multiprotein complex on a consensus SCL binding site. Forced expression of Ldb1 inhibited erythroid differentiation in G1ER proerythroblast cells, indicating Ldb1/LMO2 maintains erythroid precursors in an immature state. |
Co-immunoprecipitation, pull-down assays, forced expression/overexpression in erythroid cell lines with differentiation readout |
Proceedings of the National Academy of Sciences of the United States of America |
High |
9391090
|
| 1998 |
NLI/LDB1 mediates homo- and heteromeric complex formation between LIM-domain transcription factors (e.g., Isl1, Isl2, Lhx1, Lhx3), requiring both the N-terminal dimerization domain and C-terminal LIM interaction domain of NLI. NLI disrupts direct LIM–homeodomain interactions between Lhx3 and Isl1/Isl2, demonstrating that NLI modifies the conformational state of LIM-HD complexes. |
Co-immunoprecipitation, in vitro pull-down, yeast two-hybrid |
The Journal of biological chemistry |
High |
9452425
|
| 1998 |
Ldb1 binds LIM domains through its C-terminal region and dimerizes via its N-terminal region. Optimal Ldb1 binding requires tandem LIM domains, although single LIM domains can bind at lower efficiency. Both the dimerization and LIM-binding domains of Ldb1 are required for synergistic activation of downstream genes in Xenopus animal explant experiments. |
In vitro binding assays with deletion constructs, Xenopus animal cap explant assay |
The Journal of biological chemistry |
High |
9468533
|
| 1998 |
LDB1 (NLI) interacts with LMO1 in a T-cell leukemia line; LDB1 is a phosphoprotein and binds LMO1 in its phosphorylated state, with essentially all LMO1 and LDB1 protein in the cell forming a complex. This interaction is implicated in LMO1-driven tumorigenesis after chromosomal translocation. |
Co-immunoprecipitation from T-cell line lysates, phosphorylation analysis |
Oncogene |
Medium |
9872335
|
| 1998 |
LMO4 was identified as a novel binding partner of LDB1/NLI1 via its LIM domains. The LDB1–LMO4 interaction was characterized, and displacement of LMO4 from LDB1 complexes by ectopically expressed LMO1 or LMO2 was proposed as a mechanism contributing to T-cell leukemia. |
Yeast two-hybrid screening and co-immunoprecipitation |
Oncogene |
Medium |
9840944
|
| 2002 |
Ssdp proteins (single-stranded DNA-binding proteins) are components of Ldb1-associated nuclear complexes in HeLa cells and multiple mammalian cell types. The association is specific and does not depend on nucleic acids. Ssdp functionally cooperates with Ldb1 in Xenopus axis induction (with Xlim1), and Ssdp interacts with Chip (Drosophila Ldb1 ortholog) in wing development, demonstrating evolutionarily conserved cofactor function. |
Co-immunoprecipitation, pull-down assays, Xenopus microinjection, Drosophila genetic interaction |
Proceedings of the National Academy of Sciences of the United States of America |
High |
12381786
|
| 2003 |
High-resolution X-ray crystal structures of LMO2 and LMO4 N-terminal LIM domains in complex with the Ldb1-LID were solved (PDB: 1M3V and 1J2O). Ldb1-LID binds in an extended conformation, contributing a third strand to a beta-hairpin in the LIM1 domain. This constitutes the first molecular definition of LIM-mediated protein–protein interactions. |
NMR solution structure determination |
The EMBO journal |
High |
12727888
|
| 2003 |
Targeted deletion of Ldb1 in mice causes early embryonic lethality with no heart anlage, truncated anterior head structures, posterior axis duplication in ~40% of mutants, and severe defects in mesoderm-derived extraembryonic structures. Abnormal organizer gene expression during gastrulation and curtailed expression of Wnt inhibitors underlie axis defects. |
Gene targeting / knockout mouse, histological and molecular analysis |
Development (Cambridge, England) |
High |
12490556
|
| 2004 |
X-ray crystal structure of LMO4 in complex with Ldb1-LID was solved at high resolution. The complex has a highly modular structure with Ldb1-LID binding in an extended manner across both LIM domains of LMO4, with extensive hydrophobic/electrostatic interactions and multiple backbone hydrogen bonds. Mutagenic screen of Ldb1-LID by yeast two-hybrid and competition ELISA identified key interface residues, showing the interaction is tolerant to mutation. |
X-ray crystallography, mutagenesis, yeast two-hybrid, competition ELISA |
The EMBO journal |
High |
15343268
|
| 2005 |
Ssdp1 (encoded by headshrinker locus) regulates head morphogenesis by activating Lim1-Ldb1 transcriptional complexes. Ssdp1 contains a transcriptional activation domain, enhances Lim1-Ldb1-driven transcription in transfected cells, and interacts genetically with Lim1 and Ldb1 in head development and body growth. |
Mouse mutant analysis, transfection reporter assays, genetic interaction (compound mutant analysis) |
Development (Cambridge, England) |
High |
15857913
|
| 2006 |
The key LMO2-binding determinants in Ldb1 were mapped: both LIM domains of LMO2 are required for high-affinity Ldb1 binding (KD ~20 nM); LIM1 alone mediates primary contact, LIM2 increases affinity ~10-fold. Alanine scanning of Ldb1-LID and phage display identified 'hot spot' residues in the LIM1-binding region. LMO4 binds Ldb1 with ~2-fold higher affinity than LMO2. |
ELISA, yeast two-hybrid, phage display, alanine scanning mutagenesis |
Journal of molecular biology |
High |
16616188
|
| 2006 |
Novel Ldb1-interacting proteins in erythroleukaemic cells were identified: the repressor Eto-2 (and Mtgr1), the CDK9 kinase, and the bridging factor Lmo4. Morpholino-mediated knockdown in zebrafish showed these factors are essential for definitive haematopoiesis. Change in subcellular localization of Eto-2 was linked to transition from haematopoietic progenitor expansion to stem cell establishment. |
Mass spectrometry interactome, co-immunoprecipitation, morpholino knockdown in zebrafish, subcellular localization analysis |
Development (Cambridge, England) |
High |
17108004
|
| 2007 |
NLI/Ldb1 and erythroid partners GATA-1, SCL, LMO2 bind the β-globin locus control region (LCR) in vivo. The C-terminal LIM interaction domain of NLI is required for chromatin complex formation. Loss of this domain converts NLI into a dominant-negative inhibitor of globin expression. NLI knockdown (shRNA) prevents β-globin activation. Kinetic studies show the NLI complex appears at the β-globin promoter coincident with RNA Pol II recruitment and chromatin loop formation during erythroid differentiation. |
ChIP, shRNA knockdown, dominant-negative overexpression, chromatin conformation capture (3C) |
Molecular cell |
High |
18082606
|
| 2008 |
TAL1, LMO2, Ldb1, and E12 assemble into a five-component complex (with DNA). TAL1/E12 bHLH heterodimers form preferentially over homodimers; LMO2 binds the TAL1/E12 heterodimer with high affinity (~10^8 M^-1). The TAL1/E12/LMO2 complex forms in the presence or absence of DNA, but different complexes preferentially bind different E-box sequences. |
Analytical ultracentrifugation, isothermal titration calorimetry, fluorescence anisotropy, EMSA |
Proteins |
High |
17910069
|
| 2009 |
Ldb1 (and Ldb2) interact directly with the Ste20-like kinase SLK through the SLK C-terminal AT1-46 homology domain, in vitro and in vivo. Ldb1 and SLK colocalize in migrating cells. Both knockdown and overexpression of Ldb1 increase cell motility and focal adhesion turnover in fibroblasts, indicating Ldb1 maintains SLK in an inactive state to regulate cell migration. |
Co-immunoprecipitation, in vitro pull-down, siRNA knockdown, live-cell migration assays, confocal colocalization |
Molecular biology of the cell |
Medium |
19675209
|
| 2010 |
The Ldb1 complex (with Gata1/Tal1) acts almost exclusively as a transcriptional activator genome-wide in erythroid cells, binding a specific combination of sequences. Activation is accompanied by net decrease in binding of negative regulators Eto2 and Mtgr1. Chromosome Conformation Capture sequencing (3C-seq) demonstrated that Ldb1 complex binding marks genomic interaction sites in vivo. |
ChIP-seq, 3C-seq, genome-wide binding analysis |
Genes & development |
High |
20123907
|
| 2010 |
Ldb1 controls β-globin gene expression at multiple levels: (1) stabilizes erythroid complex partners (SCL, GATA-1, LMO2) on β-globin chromatin even though it is not itself a DNA-binding component; (2) is required for enrichment of P-TEFb (which phosphorylates RNA Pol II CTD Ser2 for elongation) at the locus; (3) is necessary for migration of the locus away from the nuclear periphery to nuclear transcription factories. |
shRNA knockdown, ChIP, chromosome conformation capture, nuclear localization imaging |
Blood |
High |
20570862
|
| 2010 |
SSBP2 stabilizes LDB1 from proteasomal degradation. Loss of Ssbp2 in mice causes increased LDB1 turnover in the thymus. SSBP2-regulated LDB1 stability controls expression of pTalpha, a target of LDB1-containing complexes critical for T-cell differentiation. |
Gene targeting (Ssbp2 knockout mice), protein turnover analysis, target gene expression |
Oncogene |
High |
20348955
|
| 2011 |
Alternative NLI (Ldb1 human homolog) complexes mediate γ-globin transcription or silencing in human erythroid cells through long-range LCR interactions. In β-globin-transcribing cells, NLI core complex at BGL3 includes corepressor ETO2 and BCL11A and the LCR contacts the β-globin gene. When γ-globin is reactivated, ETO2 participation diminishes, BCL11A occupancy is reduced, and LCR contacts shift to the BGL3/γ-globin region. |
ChIP, chromosome conformation capture (3C), RNA analysis in human erythroid cells |
Blood |
High |
22010104
|
| 2012 |
The LDB1 dimerization domain (DD) is necessary and, when fused to LMO2, sufficient to completely restore LCR-promoter looping and transcription in LDB1-depleted erythroid cells. A conserved helical region (DD4/5) within the DD is dispensable for LDB1 dimerization and chromatin looping but essential for transcriptional activation; DD4/5 recruits coregulators FOG1 and the NuRD complex, and its absence alters histone acetylation, RNA Pol II recruitment, and nuclear migration of the locus. |
shRNA depletion, domain mutant rescue, chromosome conformation capture, ChIP, nuclear localization imaging |
Genes & development |
High |
24874989
|
| 2012 |
Solution structure of the Ldb1(LID)–Lhx3 complex determined by SAXS confirms the NMR structure as an ensemble with two well-defined halves (each comprising a LIM domain from Lhx3 and a binding motif in Ldb1) with flexibility between them. A CPHDS-causing Lhx3 mutation (Y114C) does not alter zinc ligation but causes structural rearrangement of the LIM2 hydrophobic core, destabilizing the domain and reducing affinity for both Ldb1 and Isl1. |
NMR, small-angle X-ray scattering (SAXS), mutagenesis |
PloS one |
High |
22848397
|
| 2014 |
LDB1-mediated enhancer looping can be established independently of mediator and cohesin in erythroid cells. CRISPR deletion of the β-globin TATA-box eliminated transcription but not LCR/β-globin proximity. Deletion of the GATA1 site eliminated LDB1 and mediator occupancy and abrogated looping. Expression of a looping-competent but transcription-deficient LDB1 restored LCR proximity without mediator core occupancy. Cas9-directed tethering of mutant LDB1 to the promoter forced LCR loop formation in the absence of mediator or cohesin. |
CRISPR/Cas9 genome editing, Cas9-mediated protein tethering, ChIP, 3C |
Nucleic acids research |
High |
28520978
|
| 2015 |
The Isl1/Ldb1 complex promotes long-range enhancer-promoter interactions at Mef2c and Hand2 loci in cardiac progenitor cells. Ldb1 binds Isl1 and protects it from proteasomal degradation. Chromosome conformation capture sequencing identified specific Ldb1-mediated interactions of the Isl1/Ldb1-responsive Mef2c anterior heart field enhancer with cardiac progenitor genes; Ldb1 depletion downregulates these genes. |
Co-immunoprecipitation, ChIP-seq, 4C-seq, shRNA knockdown, protein stability assays |
Cell stem cell |
High |
26321200
|
| 2015 |
LDB1 mediates enhancer looping and regulates transcription in corticotrope pituitary cells via interaction with the enhancer-binding protein ASCL1. LDB1-dependent looping activates genes at the level of transcriptional initiation. For repressed genes, LDB1 looping delivers MTA2 (a NuRD complex component) to promoters to enforce promoter pausing as the repression mechanism. |
ChIP-seq, chromatin conformation capture, siRNA knockdown, RNA Pol II pausing analysis |
Proceedings of the National Academy of Sciences of the United States of America |
High |
25605944
|
| 2015 |
LDB1 is required for maintenance of fetal and adult mouse hematopoietic stem cells (HSCs). Deletion of Ldb1 in hematopoietic progenitors downregulates many transcripts required for HSC maintenance. ChIP-seq identified Ldb1 complex-binding sites at conserved regions in promoters of HSC maintenance genes, establishing Ldb1 as a central transcriptional regulator of HSC homeostasis. |
Conditional knockout mice (Cre-lox), ChIP-seq, transcriptomic profiling |
Nature immunology |
High |
21186366
|
| 2015 |
Alanine scanning mutagenesis of the LDB1 LIM interaction domain (LID) identified the motif R(320)LITR as required for LMO2 binding. Wild-type LDB1 coexpression increased LMO2 steady-state abundance; LDB1 mutants deficient in LMO2 binding compromised LMO2 stability. Mass spectrometric analysis confirmed that LMO2/LDB1 function in multisubunit complexes in leukemic cells that protect LMO2 from degradation. |
Alanine scanning mutagenesis, co-immunoprecipitation, protein stability assays (pulse-chase), mass spectrometry |
Molecular and cellular biology |
High |
26598604
|
| 2017 |
The LDB1 complex co-opts CTCF for erythroid lineage-specific long-range enhancer interactions. An LDB1-bound enhancer upstream of Car2 activates Car2 expression by looping directly to CTCF at the Car2 promoter. Both LDB1 and CTCF are required for enhancer-Car2 looping. The domain of LDB1 contacted by CTCF is necessary to rescue Car2 transcription in LDB1-deficient cells. Genome-wide studies and CRISPR/Cas9 editing indicate that LDB1–CTCF enhancer looping activates a substantial fraction of erythroid genes. |
ChIP, chromosome conformation capture, CRISPR/Cas9 genome editing, LDB1 domain rescue assays |
Cell reports |
High |
28636938
|
| 2019 |
Crystal structure of human LDB1 in complex with SSBP2 at 2.8-Å resolution revealed: (1) the LDB1 dimerization domain contains an NTF2-like subdomain and a small helix4–helix5 (H4-H5) subdomain forming the dimerization interface; (2) two LDB/Chip conserved domains (LCCDs) flank the core DDs in the symmetric dimer; (3) each LCCD forms extensive interactions with an SSBP2 dimer; (4) a conserved linker between LDB1 DD and LCCD covers a potential ligand-binding pocket of the NTF2-like subdomain. |
X-ray crystallography at 2.8 Å, biochemical validation |
Proceedings of the National Academy of Sciences of the United States of America |
High |
31892537
|
| 2019 |
LHX2 and LDB1 regulate the assembly and maintenance of interchromosomal olfactory receptor (OR) compartments and Greek island (OR enhancer) hubs in olfactory sensory neurons. In situ Hi-C on sorted neurons showed Greek islands from 18 chromosomes form specific interchromosomal contacts increasing with differentiation. Loss of LHX2 or LDB1 disrupts these trans interactions and OR transcription. |
In situ Hi-C on FACS-sorted neurons, LHX2/LDB1 conditional knockout, ChIP |
Nature |
High |
30626972
|
| 2019 |
Ldb1 is recruited to Pax3-bound chromatin elements independently of CTCF-Cohesin in myogenic progenitors and mediates long-range chromatin looping interactions at a subset of Pax3 binding sites, promoting H3K4me1 deposition and looping. Ldb1 deletion in Pax3-expressing cells in vivo severely impairs specification of migratory myogenic progenitors. |
Mass spectrometry, ChIP-seq, chromosome conformation capture, conditional knockout mice |
Nature communications |
High |
31127120
|
| 2020 |
LDB1 confers enhanced protein stability on direct binding partners (LMO2, SSBP) and indirect partners (TAL1) in leukemic cells. Protein half-lives are ordered: LDB1 > SSBP > LMO2 > TAL1. LDB1 is a remarkably stable scaffold protein that nucleates multisubunit complex assembly; free subunits are more rapidly degraded than those incorporated in the LMO2/LDB1 complex. |
Halo protein tagging, pulse-chase/kinetic stability assays, variant proteins deficient in partner binding |
Molecular and cellular biology |
High |
32229578
|
| 2020 |
Ldb1 is required for Lmo2-induced thymocyte self-renewal, thymocyte radiation resistance, and transition from preleukemic thymocytes to T-ALL. Co-binding of Ldb1 and Lmo2 was detected at promoters of key T-ALL driver genes (Hhex, Lyl1, Nfe2), and Cre-mediated Ldb1 deletion reduced binding of both Ldb1 and Lmo2 at these sites, establishing that the Ldb1/Lmo2 complex directly drives the leukemic transcriptional program. |
Conditional Ldb1 knockout in Lmo2 transgenic T-ALL mouse model, ChIP-seq, transcriptomic profiling |
Blood |
High |
32181817
|
| 2020 |
The Lhx2–Ldb1 complex regulates hippocampal cell fate specification in an evolutionarily conserved manner. A chimeric construct encoding the Lhx2 homeodomain fused to the Ldb1 dimerization domain cell-autonomously rescues comprehensive hippocampal deficits (field-specific molecular identity, neuron-glia cell fate switch) in Ldb1 conditional mutant mice, demonstrating the LHX2:LDB1 tetrameric complex as the essential molecular device. |
Conditional knockout mice, in utero electroporation of rescue construct, molecular phenotyping |
Development (Cambridge, England) |
High |
32994168
|
| 2025 |
SSBP3 (and SSBPs broadly) are essential functional components of the architectural LDB1 complex. SSBP3 is essential for murine erythroid cell viability, LDB1 function, and transcription. LDB1, not single-stranded DNA, is the predominant genome-wide chromatin tether of SSBP3. SSBP3 depletion in SSBP2/4 knockout cells globally weakens LDB1-dependent chromatin loops and reduces nascent transcription without affecting LDB1's chromatin binding. Chromatin tethering experiments show SSBP3 and LDB1 mutually depend on each other for looped contacts. In vitro biochemistry shows SSBP3 stabilizes LDB1 dimerization. |
CRISPR knockout, genome-wide ChIP-seq, Hi-ChIP, SSBP2/4 double knockout combined with SSBP3 depletion, chromatin tethering, in vitro dimerization assay |
Molecular cell |
High |
40803327
|
| 2025 |
Drosophila Ldb1 (Chip) functions as a cofactor of the pioneer factor Zelda during zygotic genome activation. Rapid nuclear depletion (optogenetics/iLEXY) showed Chip's essential function is limited to a 1-hour window overlapping ZGA. Zelda recruits Chip to chromatin; Chip does not significantly impact chromatin architecture at these stages but recruits CBP and is essential for H3K27ac deposition at enhancers and promoters and for co-regulated gene expression. |
Optogenetic rapid nuclear depletion (iLEXY), ChIP-seq, ATAC-seq, CUT&RUN, transcriptomics in Drosophila embryos |
Molecular cell |
High |
40494353
|
| 2021 |
FLI-1 interacts with the LDB1 complex and enables recruitment of the LDB1 complex to regulatory sequences of megakaryocytic genes and enhancers. FLI-1 promotes chromatin looping between enhancers and promoters through the LDB1 complex during megakaryopoiesis. |
Co-immunoprecipitation, ChIP-seq, chromosome conformation capture, shRNA knockdown |
iScience |
Medium |
33733070
|
| 2022 |
LDB1 interacts with BLIMP1/PRDM1 (in C. elegans, LDB-1 with BLMP-1) and participates in both transcriptional activation and repression of a subset of BLIMP1-regulated genes. LDB-1 and HAM-3 (SWI/SNF subunit) bind BLMP-1; human LDB1, SMARCD3/BAF60C, and SMARCC1/BAF155 all physically interact with human BLIMP1/PRDM1 in vitro and are closely associated in vivo. |
C. elegans genetics, RNAi, co-immunoprecipitation, human in vitro pull-down |
Biochimica et biophysica acta. Gene regulatory mechanisms |
Medium |
32417234
|
| 2022 |
Cytoplasmic LMO2–LDB1 complex activates STAT3 signaling in glioma stem cells (GSCs) through interaction with gp130 and JAK1/2. LMO2-driven STAT3 phosphorylation requires LDB1 and leads to increased expression of ID1 (stemness regulator), demonstrating a non-nuclear signaling function for the LMO2–LDB1 complex. |
Co-immunoprecipitation, proximity ligation assay, LDB1/LMO2 knockdown with STAT3 phosphorylation readout, subcellular fractionation |
Cells |
Medium |
35805116
|