| 2000 |
EB3 is a novel member of the EB1 family that associates with the cytoplasmic microtubule network and binds directly to APCL (APC2), a CNS-specific homologue of APC, as demonstrated by yeast two-hybrid screening and confocal microscopy showing co-localization in perinucleus and microtubule network. |
Yeast two-hybrid, confocal microscopy, sequence analysis |
Oncogene |
Medium |
10644998
|
| 2003 |
EB3-GFP tracks exclusively to the growing plus-ends of microtubules in live neurons, marking microtubule growth events; microtubules grow slower in neurons than in glia/COS-1 cells, and ~65% of EB3-GFP movements in proximal dendrites are directed distally vs. ~35% toward the cell body, reflecting the mixed microtubule polarity of dendrites. |
Live-cell fluorescence microscopy with EB3-GFP in cultured neurons |
The Journal of neuroscience |
High |
12684451
|
| 2005 |
EB1 and EB3 directly bind CLIP-170 and CLIP-115 through their C-terminal tyrosine residues, and control CLIP dissociation kinetics from microtubule plus-ends; RNAi depletion of EB1 and EB3 accelerates CLIP dissociation from tips, which is rescued by EB1 but not EB2 expression. |
RNA interference, direct binding assay, live-cell imaging of CLIP dynamics |
Molecular biology of the cell |
High |
16148041
|
| 2007 |
EB3 is specifically upregulated upon myogenic differentiation; knockdown of EB3 (but not EB1) prevents myoblast elongation and fusion, impairs microtubule capture at the cell cortex, and disrupts microtubule dynamics; two specific amino acids in the calponin-like domain of EB3 are required for myoblast fusion. |
RNAi knockdown, live-cell imaging, EB1/EB3 chimera expression, C2C12 myoblast differentiation assay |
Current biology : CB |
High |
17658256
|
| 2007 |
EB3 interacts with the p53-target DDA3 protein; the interaction requires intact microtubules, maps to aa 118-329 of DDA3 and both N- and C-termini of EB3; DDA3 and EB3 cooperate for microtubule binding in vitro; ectopic expression of DDA3 and EB3 enhances beta-catenin-dependent transactivation and cyclin D1 production. |
Yeast two-hybrid, GST pull-down, co-immunoprecipitation, in vitro microtubule-binding assay, immunofluorescence, reporter assay |
Oncogene |
Medium |
17310996
|
| 2008 |
EB3 directly binds drebrin (an F-actin-associated protein); in growth cones this interaction occurs specifically when drebrin is on F-actin in the proximal region of filopodia and EB3 is at microtubule tips invading filopodia; disruption of this interaction impairs growth cone formation and neurite extension. |
Direct binding assay (in vitro), co-localization, dominant-negative disruption, neurite outgrowth assay |
Nature cell biology |
High |
18806788
|
| 2009 |
EB3 stability is regulated during mitosis by Aurora-A and Aurora-B kinases that phosphorylate EB3 at Ser-176, disrupting the EB3-SIAH-1 complex; SIAH-1 ubiquitin ligase mediates EB3 polyubiquitination and proteasomal degradation during G1; phosphorylation-induced stabilization of EB3 during mitosis facilitates cell cycle progression at prometaphase. |
In vitro kinase assay, co-immunoprecipitation, RNAi knockdown, proteasome inhibition, cell cycle analysis |
The Journal of biological chemistry |
High |
19696028
|
| 2010 |
In vitro reconstitution demonstrated that EB3 restores robust microtubule growth in the presence of MCAK (which alone blocks assembly) and targets MCAK to growing microtubule ends by increasing its association rate; the EB3-dependent targeting requires direct EB3-MCAK interaction and enhances MCAK's capacity to induce catastrophes without affecting growth/shortening velocities. |
In vitro reconstitution of microtubule dynamics, TIRF microscopy, quantitative analysis of dynamics |
Current biology : CB |
High |
20850319
|
| 2011 |
EB3 (and EB1) directly interact with the AIS scaffold protein ankyrin G (ankG); EB3 is concentrated and stabilized at the axon initial segment (AIS) in mature neurons; knockdown of ankG leads to cell-wide upregulation of EB3 comets; EB3/EB1 participate in AIS maintenance. |
Direct interaction assay, live-cell imaging, immunofluorescence, ankG knockdown, fractionation/localization |
Proceedings of the National Academy of Sciences of the United States of America |
High |
21551097
|
| 2011 |
EB3 (and EB1) are required for assembly of primary cilia; cells lacking EB1 or EB3 have defective MT minus-end anchoring at centrosome/basal body and short cilia stumps; GST pull-down and mass spectrometry showed EB1/EB3 interact with proteins implicated in MT minus-end anchoring and vesicular trafficking to cilia base; EB3 localizes to the tip of motile cilia and affects centriole-associated rootlet filament formation. |
Protein depletion (siRNA), dominant-negative expression, electron microscopy, GST pull-down, mass spectrometry, immunoprecipitation, live imaging |
Journal of cell science |
High |
21768326
|
| 2011 |
The SH3 domain of PSD-95 interacts with a proline-rich region within EB3; overexpression of PSD-95 decreases the lifetime of EB3 comets in dendrites, leading to less organized microtubules at dendritic branch points and decreased dendritic branching. |
Co-immunoprecipitation, live-cell imaging (EB3 comet analysis), overexpression in neurons |
The Journal of neuroscience |
Medium |
21248129
|
| 2012 |
VE-cadherin outside-in signaling activates Src and PLCγ2, causing Ca2+ release from ER stores, activating calcineurin (CaN); CaN downregulation leads to phosphorylation of EB3 at Ser-162, destabilizing the EB3 dimer, suppressing microtubule growth, and enabling adherens junction assembly; phospho-defective S162A EB3 mutant induces MT growth in confluent monolayers and disassembles AJs. |
Phospho-specific mutagenesis, calcium imaging, pharmacological inhibition, co-immunoprecipitation, endothelial permeability assay |
Molecular cell |
High |
23159740
|
| 2012 |
Drebrin E and EB3 form a complex with myosin IIB and βII-spectrin at the apical domain of columnar epithelial cells; depletion of drebrin E disrupts apical accumulation of EB3 and impairs cell elongation; EB3 depletion produces a similar elongation defect; the complex connects F-actin and microtubule networks apically during epithelial morphogenesis. |
Co-immunoprecipitation, siRNA knockdown, immunofluorescence, morphometric analysis |
Journal of cell science |
Medium |
22275434
|
| 2013 |
Aurora B phosphorylates EB3 at Ser-176 at the midbody to control cortical microtubule growth; EB3 stabilizes focal adhesions and coordinates daughter cell spreading during mitotic exit, promotes midbody microtubule stability, and is required for efficient cytokinesis; EB1 and EB3 play temporally distinct roles in cell division, with EB1 involved in spindle orientation before anaphase. |
Phospho-specific mutagenesis, live-cell imaging, RNAi knockdown, immunofluorescence |
The Journal of cell biology |
High |
23712260
|
| 2014 |
BPAG1a/b C-terminal isoform-specific tails bind both EB1 and EB3 and are sufficient to bundle microtubules; knockdown of BPAG1a/b in C2.7 myoblasts impairs directness of cell migration and disrupts Golgi structure. |
GST pull-down, co-immunoprecipitation, microtubule bundling assay, siRNA knockdown, migration assay |
PloS one |
Medium |
25244344
|
| 2015 |
EB3 binds to IP3 receptors (IP3R3) through an S/TxIP EB-binding motif; in endothelial cells, EB3 depletion or mutation of the TxIP motif of IP3R3 prevents α-thrombin-induced IP3R3 clustering, Ca2+ increase, myosin light chain phosphorylation, and vascular permeability increase; selective EB3 gene deletion in mouse endothelial cells abrogates α-thrombin-induced endothelial permeability. |
Co-immunoprecipitation, mutagenesis of binding motif, siRNA knockdown, conditional knockout mouse, Ca2+ imaging, permeability assay |
Cell reports |
High |
26119739
|
| 2017 |
EB3 (and EB1) form a complex with myomegalin that acts as a membrane-microtubule tether at Golgi membranes; CRISPR/Cas9 knockout of EB2/EB3 and C-terminal half of EB1 reduces CAMSAP2-decorated microtubule minus end lengths, detaches microtubules from Golgi membranes, compacts the Golgi complex, and disrupts cell migration, polarity, and focal adhesion distribution. |
CRISPR/Cas9 knockout, co-immunoprecipitation, electron microscopy, live-cell imaging, immunofluorescence |
The Journal of cell biology |
High |
28814570
|
| 2017 |
STIM2 forms a Ca2+-dependent complex with EB3 via a Ser-x-Ile-Pro (SxIP) amino acid motif; disruption of STIM2-EB3 interaction results in loss of mushroom dendritic spines; EB3 overexpression rescues mushroom spine loss in a PS1-M146V knock-in Alzheimer's disease model, while STIM2 overexpression fails to rescue spines after EB3 knockdown. |
Co-immunoprecipitation, mutagenesis of SxIP motif, siRNA knockdown, overexpression, spine morphometry in hippocampal neurons |
Scientific reports |
Medium |
29247211
|
| 2018 |
IP3K-A (ITPKA) binds to EB3, and this interaction is regulated by PKA-dependent phosphorylation of IP3K-A at Ser119; the IP3K-A/EB3 complex dissociates and reassociates rapidly during chemically-induced LTP conditions. |
Co-immunoprecipitation, phospho-specific mutagenesis, chemical LTP induction |
Biochemical and biophysical research communications |
Low |
30466786
|
| 2019 |
EB3 comet intensity profiles in mouse fibroblasts frequently exhibit secondary EB3-binding peaks (EB3-islands) on 56% of growing microtubule comets, which are stationary and originate from EB3 comets moving with growing MT tips; modeling suggests additional protein factors beyond GTP hydrolysis contribute to EB3 residence time on MTs. |
Live-cell fluorescence microscopy (EB3-RFP), quantitative image analysis, computational modeling |
Molecular biology of the cell |
Medium |
31141458
|
| 2021 |
Loss of EB3 (but not EB1) inhibits neuritogenesis in embryonic cortical neurons; EB3 is more proximally located at dynamic microtubule plus-ends than EB1 in growth cone filopodia, enabling continuous microtubule elongation as the drebrin/EB3 pathway zippers microtubules to F-actin; pharmacological blockade of microtubule dynamics is associated with selective loss of EB3 (not EB1) from plus-ends. |
Gene editing (CRISPR), pharmacological inhibition, overexpression, live-cell imaging, morphometric analysis |
Journal of neurochemistry |
High |
34478582
|
| 2023 |
Fidgetin (microtubule-severing enzyme) trims tyrosinated (labile) microtubules by interacting with EB3; fidgetin depletion increases EB3 at neurite ends, elongates the labile microtubule domain, and increases axon length and branching; EB3 knockdown does not change fidgetin expression but fidgetin deletion markedly increases EB3 levels. |
RNA interference, overexpression, rat SCI model, immunofluorescence, EB3 localization analysis |
Neural regeneration research |
Medium |
37449637
|
| 2023 |
A 14-amino acid peptide (CIPRI) targeting the EB3-IP3R3 interface disrupts EB3-IP3R3 interaction in vitro and in mouse lungs; disruption mitigates Ca2+ release from ER stores, prevents VE-cadherin junction disassembly, and reduces inflammation-induced lung injury and microvascular leakage in mice. |
Peptide competition assay, co-immunoprecipitation, Ca2+ imaging, in vivo endotoxemia/sepsis mouse model, vascular permeability assay |
American journal of respiratory cell and molecular biology |
High |
37290041
|
| 2024 |
EZH2-mediated H3K27 trimethylation epigenetically represses MAPRE3 (EB3) expression at its promoter in ovarian cancer; EZH2 knockdown decreases H3K27me3 enrichment at the MAPRE3 promoter and increases EB3 expression; MAPRE3 overexpression induces cell cycle arrest and apoptosis in ovarian cancer cells. |
ChIP assay (H3K27me3 at MAPRE3 promoter), EZH2 knockdown, overexpression, xenograft tumor model |
Experimental cell research |
Medium |
38199479
|
| 2024 |
EB3 undergoes liquid-liquid phase separation (LLPS) with significantly higher propensity than EB1 despite 67% sequence identity; this difference involves contributions from multiple protein regions with histidine residues in the N-terminal domain playing a key role; EB3 condensates have lower dynamics (less fluid) than EB1 and higher capacity to recruit tubulin and nucleate polymerization; EB3/CLIP-170 droplets show higher tubulin polymerization capacity than EB1/CLIP-170. |
In vitro LLPS assay, fluorescence recovery after photobleaching (FRAP), mutagenesis, in vitro tubulin polymerization assay, cell-based condensate imaging |
The Journal of biological chemistry |
High |
41135671
|
| 2024 |
In vitro reconstitution showed that EB3-dependent comet size at microtubule ends is independent of GTP-cap size; a phenomenological model based on noisy microtubule growth and a single EB3-dependent hydrolysis rate fully describes stalled and freely growing microtubule lifetime distributions; modulation of microtubule growth velocity in cells does not consistently alter EB3 comet length. |
In vitro reconstitution with microfabricated barriers, GFP-EB3 live imaging, mathematical modeling |
Biophysical journal |
Medium |
39604262
|