| 2003 |
Tuba (DNMBP) is a scaffold protein containing four N-terminal SH3 domains that bind dynamin, a central DH domain that functions as a Cdc42-specific GEF (not activating Rac or Rho), a BAR domain (which replaces the typical PH domain following DH domains), and a C-terminal SH3 domain that directly binds N-WASP and Ena/VASP proteins, linking dynamin to actin regulatory proteins. |
Co-immunoprecipitation, pulldown assays, in vitro GEF activity assay, forced mitochondrial targeting of C-terminal SH3 domain to assess F-actin recruitment |
The Journal of biological chemistry |
High |
14506234
|
| 2005 |
The four N-terminal SH3 domains of Tuba bind dynamin with exceptionally high avidity; the DH domain is a Cdc42-specific GEF unique in being followed by a BAR domain rather than a PH domain; the C-terminal SH3 domain directly binds N-WASP and Ena/VASP and recruits a larger actin regulatory complex. |
Biochemical pulldown, co-immunoprecipitation, GEF activity assays, domain characterization |
Methods in enzymology |
High |
16413298
|
| 2006 |
Tuba is concentrated at the apical-most region of cell junctions in simple epithelia via interaction with ZO-1. RNAi-mediated depletion of Tuba alters the geometrical configuration of cell junctions (curved/slack appearance), modifies junctional F-actin and E-cadherin assembly, and retards junction formation. Suppression of Cdc42 or N-WASP mimics Tuba depletion; overexpression of dominant-active Cdc42 or N-WASP rescues Tuba-depleted cells, establishing a Tuba→Cdc42→N-WASP pathway for junction shaping. |
RNAi knockdown, immunofluorescence, calcium-switch junction formation assay, dominant-active rescue, co-immunoprecipitation with ZO-1 |
The Journal of cell biology |
High |
17015620
|
| 2006 |
Tuba overexpression stimulates dorsal ruffles and actin-driven motility of intracellular puncta requiring the C-terminal SH3, DH/GEF, and BAR domains. Tuba is recruited to PIP5Kα-generated lipid vesicles and promotes N-WASP-dependent actin comet formation. RNAi knockdown of Tuba attenuates PIP5Kα-generated comet formation and invasive behavior of B16 melanoma cells. |
Overexpression, RNAi knockdown, fluorescence microscopy, domain deletion analysis, invasiveness assays |
Journal of cell science |
Medium |
16757518
|
| 2010 |
Tuba, as a Cdc42-specific GEF, is required for Cdc42 enrichment at the apical cortex of epithelial cysts. Loss of Tuba causes a multilumen phenotype by disrupting spindle orientation; this is rescued by human Tuba or constitutively active Cdc42 but not by a GEF-dead Tuba mutant, placing Tuba upstream of Cdc42→aPKC→spindle orientation in epithelial ductal morphogenesis. |
RNAi screen (70 GEFs), RNAi knockdown, rescue with Tuba WT vs. GEF-dead mutant, active Cdc42 rescue, immunofluorescence, aPKC activity assay |
The Journal of cell biology |
High |
20479467
|
| 2011 |
Tuba and N-WASP function cooperatively at the pre-apical patch (PAP) to position the central lumen during epithelial cyst morphogenesis; each depends on the other for localization to the PAP, and the cooperative function requires the polyproline region of N-WASP. |
RNAi knockdown, immunofluorescence localization, multilumen phenotype assay in Caco-2 cysts |
Cell adhesion & migration |
Medium |
21677511
|
| 2013 |
The Listeria virulence factor InlC binds the sixth SH3 domain (SH3-6) of human Tuba, competitively disrupting its physiological interaction with N-WASP and Mena; the InlC/Tuba SH3-6 interaction is centered on Phe146 of InlC stacking on Asn1569 of Tuba, and replacing Phe146 with Ala largely abrogates affinity and in vivo mimics deletion of inlC. |
Crystal structure of Tuba SH3-6 complexes with InlC, N-WASP, and Mena; site-directed mutagenesis (F146A); in vivo spreading assays |
Structure (London, England : 1993) |
High |
24332715
|
| 2014 |
DDR1 (Discoidin Domain Receptor 1) signals through Tuba and Cdc42 to promote linear invadosome formation and matrix degradation on collagen I fibrils; DDR1 kinase activity and Src are not required, but Cdc42 activation by DDR1 is Tuba-dependent, as both Tuba and Cdc42 localize to linear invadosomes and are required for their formation. |
SiRNA knockdown, immunofluorescence colocalization, Cdc42 activity assay (pulldown), matrix degradation assay, collagen gel invasion assay |
The Journal of cell biology |
High |
25422375
|
| 2016 |
Tuba (Dynamin Binding Protein) is required for ciliogenesis and nephrogenesis: Tuba knockdown in MDCK cells abolishes primary cilia, impairs apical polarization, and inhibits HGF-induced tubulogenesis. In zebrafish, tuba morphants exhibit ciliary mutant phenotypes in multiple ciliated organs; co-injection of sub-threshold tuba and cdc42 morpholinos causes genetic synergy and disorganized pronephric duct cilia, placing Tuba in the same Cdc42/ciliogenesis pathway. |
RNAi knockdown in MDCK cells, zebrafish morpholino knockdown, genetic epistasis (tuba + cdc42 double morpholino), immunofluorescence, electroretinography |
The Journal of biological chemistry |
High |
26895965
|
| 2018 |
Bi-allelic loss-of-function variants in DNMBP cause infantile cataracts in humans. RNAi knockdown of the Drosophila ortholog still life (sif) in lens-secreting cells disrupts lens-secreting cell development, delocalizes E-cadherin, alters septate junction distribution in cone cells, and reduces electroretinography amplitudes. In human epithelial cells, DNMBP regulates tight junction shape and E-cadherin assembly pattern. |
Exome sequencing (human genetics), Drosophila RNAi knockdown, immunofluorescence (E-cadherin/septate junction localization), electroretinography, human cell knockdown |
American journal of human genetics |
High |
30290152
|
| 2018 |
A Tuba/Cdc42/Par6A trimeric complex is required to ensure singularity of the apical domain during enterocyte polarization. Tuba, Cdc42, and Par6A co-immunoprecipitate and partially colocalize at the apical membrane; Par6A is required to restrict Cdc42 signaling at the apical domain; rescue experiments using Par6A mutants show that the ability to form this trimeric complex correlates with restoration of apical domain singularity. |
Co-immunoprecipitation, CRISPR knockout of Par6A, rescue with Par6A mutants, live imaging of active Cdc42 (FRAP), immunofluorescence colocalization |
PloS one |
High |
30408122
|
| 2021 |
Tuba activates Cdc42 downstream of Rab8a during neuronal polarization. Rab8a activity generates a proximal-to-distal axonal gradient of Tuba in cultured neurons; gain-of-function of Rab8a or Tuba produces supernumerary axons, while loss-of-function abrogates axon specification, phenocopying Cdc42 loss. In vivo, dominant-negative Rab8a or Tuba knockdown impairs cortical neuronal migration in mice. |
Gain-of-function and loss-of-function (RNAi/dominant negative) in cultured rat hippocampal neurons, in utero electroporation for cortical migration in mice, immunofluorescence of Tuba gradient |
The Journal of neuroscience |
High |
33478991
|
| 2025 |
The ubiquitin ligase Nedd4-2 (NEDD4L) binds DNMBP/Tuba preferentially under hyperosmotic stress and ubiquitinates it, targeting DNMBP to P-body condensates. DNMBP itself promotes P-body formation under hyperosmolarity. Both Nedd4-2 and DNMBP are required for Cdc42 activation following hyperosmotic treatment; DNMBP knockout suppresses Cdc42 and its downstream effector p38-MAPK. |
BioID proximity labeling screen (miniTurbo-Nedd4-2), co-immunoprecipitation, ubiquitination assay, P-body localization by immunofluorescence, DNMBP knockout, Cdc42 and p38-MAPK activity assays |
The Journal of biological chemistry |
Medium |
40975170
|