| 1999 |
STEF/TIAM2 is a novel guanine nucleotide exchange factor (GEF) that specifically activates Rac1 but not RhoA or Cdc42. In vitro GDP dissociation assays demonstrated selective GDP release from Rac1. The protein contains two pleckstrin homology (PH) domains, a PDZ domain, and a Dbl homology (DH) domain. Expression of a truncated STEF in cultured cells induced membrane ruffling with altered actin localization, indicating in vivo Rac1 activation. |
In vitro GDP dissociation assay, domain structure analysis, cell-based ruffling assay |
The Journal of biological chemistry |
High |
10364228
|
| 1999 |
TIAM2 (cloned independently as a TIAM1 homolog) encodes a protein with GDP-GTP exchange activity, confirmed by direct purification and exchange assay. |
Protein purification, GDP-GTP exchange assay |
Genomics |
High |
10512681
|
| 2001 |
Domain dissection of STEF revealed that the N-terminal PH domain (PHnTSS) is required for membrane association, the DH domain carries enzymatic GEF activity, and the C-terminal PH domain (PHc) promotes catalytic activity. A dominant-negative PHnTSS fragment inhibited both STEF and Tiam1 function and blocked neurite outgrowth in N1E-115 neuroblastoma cells; this inhibition was rescued by exogenous STEF or Tiam1, demonstrating STEF acts through Rac1 to drive neurite formation. |
Deletion mutagenesis, dominant-negative expression, neurite outgrowth assay, rescue experiment |
The Journal of biological chemistry |
High |
11707441
|
| 2003 |
In vivo gene transfer by in utero electroporation showed that STEF/Tiam1 (Rac1 activators) are required for radial migration of cortical neurons. Functional repression of STEF/Tiam1 or Rac1 inhibited neuronal migration and caused loss of the leading process; downstream JNK (activated by Rac1) regulates microtubule dynamics via MAP1B phosphorylation in migrating neurons. |
In utero electroporation (dominant-negative/shRNA), cortical migration assay, JNK inhibitor treatment, MAP1B phosphorylation assay |
The EMBO journal |
High |
12912917
|
| 2003 |
STEF and Tiam1 localize within growth cones of primary hippocampal neurons and are essential for formation of growth cone lamellipodia and neurite growth. STEF/Tiam1 mediate extracellular laminin signals and intracellular Cdc42 signals to activate Rac1 in the growth cone; RhoA inhibits the STEF/Tiam1-Rac1 pathway. |
Dominant-negative expression, immunolocalization in primary neurons, growth cone morphology assay, epistasis with Cdc42 and RhoA |
Molecular and cellular neurosciences |
Medium |
14550769
|
| 2005 |
PAR-3 directly interacts with STEF/Tiam1 (Rac-specific GEFs) and STEF forms a complex with PAR-3-aPKC-PAR-6-Cdc42-GTP. This complex mediates Cdc42-induced Rac activation and lamellipodia formation in neuroblastoma cells and cultured hippocampal neurons, establishing a Cdc42-PAR-6-PAR-3-STEF-Rac pathway required for neuronal polarity. Disruption of PAR-3-STEF binding inhibited Cdc42-induced lamellipodia but not filopodia. STEF accumulates at the tip of the growing axon co-localizing with PAR-3. |
Co-immunoprecipitation, direct binding assay, dominant-negative and overexpression in N1E-115 cells and hippocampal neurons, Rac1 activation assay |
Nature cell biology |
High |
15723051
|
| 2007 |
Rho-kinase phosphorylates STEF at Thr1662 in vitro, reducing STEF-induced Rac1 activation in COS7 cells. LPA-induced phosphorylation of STEF in PC12D cells is suppressed by Y-27632 (Rho-kinase inhibitor). Phosphorylation diminishes STEF interaction with microtubule-associated protein 1B (MAP1B), and a phosphomimetic STEF mutant has weakened ability to enhance NGF-induced neurite outgrowth. |
In vitro kinase assay, site-directed mutagenesis, Rac1 activation assay, Co-immunoprecipitation, neurite outgrowth assay, pharmacological inhibitor |
Biochemical and biophysical research communications |
High |
17320046
|
| 2007 |
STEF physically associates with activated Rap1 through its TSS region. Rap1-STEF interaction downstream of the cAMP/Epac/Rap1 cascade activates Rac1, which mediates non-amyloidogenic alpha-cleavage of APP (sAPPα secretion). A deleted TSS domain of STEF fails to activate Rac1 and dramatically decreases sAPPα secretion induced by Epac. |
Co-immunoprecipitation (Rap1-STEF), deletion mutagenesis (ΔTSS), Rac1 activation assay, sAPPα secretion assay |
FEBS letters |
Medium |
18047838
|
| 2010 |
STEF/TIAM2 is required for Rac activation during microtubule regrowth (following nocodazole washout), which drives focal adhesion (FA) disassembly. STEF knockdown reduces the rate of multiple FA targeting by microtubules, leading to enlarged FAs and reduced cell migration speed. |
siRNA knockdown, Rac1 activation assay (FRET/pulldown), live imaging of FA dynamics (fluorescence microscopy), nocodazole washout assay, migration speed measurement |
EMBO reports |
High |
20224579
|
| 2011 |
The PDZ domains of Tiam1 and Tiam2 have overlapping but distinct ligand specificities determined by non-conserved residues in the S(0) and S(-2) pockets. Site-directed mutagenesis of four non-conserved residues in Tiam1's PDZ domain converted its specificity to that of Tiam2, as confirmed by combinatorial peptide library screening and binding assays with native protein-derived peptides. |
Combinatorial peptide library, peptide binding assays, site-directed mutagenesis, double mutant cycle analysis |
Biochemistry |
High |
21192692
|
| 2011 |
PKA phosphorylates STEF at three residues (Thr-749, Ser-782, Ser-1562); phosphorylation at Thr-749 is specifically critical for cAMP/dbcAMP-induced Rac1 activation and neurite outgrowth in PC12D cells. STEF depletion drastically reduces dbcAMP-induced neurite outgrowth. During dbcAMP stimulation, PKA activation at the plasma membrane becomes localized to neurite tips, coinciding with local Rac1 activation via STEF. |
FRET-based biosensors (Rac1, Cdc42, PIP3), STEF siRNA knockdown, site-directed mutagenesis (phosphorylation-site mutants), neurite outgrowth assay, PKA activity reporter |
Molecular biology of the cell |
High |
21460187
|
| 2011 |
Expression of TIAM2S (short isoform) in HepG2 hepatocellular carcinoma cells promoted cell growth, invasiveness, and in vivo tumor formation in xenograft mice. TIAM2S expression upregulated N-cadherin and vimentin and caused redistribution of E-cadherin, indicating promotion of epithelial-to-mesenchymal transition (EMT). |
Stable overexpression, proliferation and invasion assays, xenograft mouse model, Western blot for EMT markers |
International journal of cancer |
Medium |
21469146
|
| 2013 |
TIAM2 knockdown by siRNA in NSCLC cells suppressed Rac1 activation (assessed by GST pulldown), reduced invasion and motility, upregulated E-cadherin, and downregulated MMP-3, Twist, and Snail, linking TIAM2-mediated Rac1 activation to EMT regulation in lung cancer cells. |
siRNA knockdown, GST-Rac1 pulldown activation assay, invasion/motility assay, Western blot for EMT markers |
Asian Pacific journal of cancer prevention |
Medium |
24377522
|
| 2018 |
STEF/TIAM2 localizes at the nuclear envelope, co-localizing with Nesprin-2G and Non-muscle myosin IIB (NMMIIB), where it regulates perinuclear Rac1 activity. STEF depletion reduces apical perinuclear actin cables (rescued by targeting active Rac1 to the nuclear envelope), increases nuclear height, impairs nuclear re-orientation, reduces perinuclear pMLC and myosin-generated tension at the nuclear envelope, decreases nuclear stiffness, and reduces TAZ-regulated gene expression. |
siRNA knockdown, fluorescence localization (co-localization with Nesprin-2G and NMMIIB), FRET-based perinuclear Rac1 biosensor, active-Rac1 nuclear envelope targeting rescue, FRAP, nuclear morphology assay, TAZ reporter |
Nature communications |
High |
29844364
|
| 2019 |
Two isoforms of TIAM2 act oppositely in regulating transmission ratio distortion (TRD) by the mouse t-haplotype: the short isoform (Tiam2s), whose expression is strongly increased from the t-allele, enhances t-haplotype transmission (sperm motility-linked), while the full-length isoform (Tiam2l) has the opposite effect. Both isoforms affect Rac1 signaling in sperm. |
Transgenic mouse approaches, transmission ratio distortion assay, isoform-specific expression analysis |
PLoS genetics |
Medium |
30817801
|
| 2021 |
TIAM2 is identified as an essential Rac-GEF responsible for Rac1-mediated lung adenocarcinoma cell migration downstream of EGFR and c-Met receptor tyrosine kinases. TIAM2 controls distinctive aspects of ruffle dynamics in a non-redundant manner with FARP1 and ARHGEF39. The AXL-Gab1-PI3K axis confers pro-motility signaling in this context. |
siRNA knockdown (functional screen), Rac1 activity assay, ruffle dynamics live imaging, epistasis with EGFR/c-Met/AXL-Gab1-PI3K pathway |
Cell reports |
Medium |
34731623
|
| 2023 |
NSUN2-catalyzed m5C methylation of TIAM2 mRNA stabilizes the transcript in a YBX1-dependent manner. Loss of NSUN2 decreases m5C modification on TIAM2 mRNA, accelerates its decay, reduces TIAM2 expression, and suppresses pancreatic cancer EMT, growth, and metastasis. Rescue experiments confirmed TIAM2 acts downstream of NSUN2. |
m5C-seq, RNA-seq, lentiviral knockdown/overexpression, mRNA stability assay, YBX1 dependency assay, in vivo xenograft |
Cell death discovery |
Medium |
37393317
|
| 2023 |
Dually lipidated CaMKIγ (prenylated and palmitoylated) functionally couples to STEF-Rac1 signaling at lipid rafts to drive neurite extension in PC12 cells. Palmitoylation is specifically required for CaMKIγ to activate the compartmentalized STEF-Rac1 pathway; prenylation alone is insufficient for this functional coupling. |
Lipid raft fractionation, palmitoylation/prenylation mutants, Rac1 activity assay, neurite outgrowth assay in PC12 cells lacking endogenous CaMKIγ |
Frontiers in cellular neuroscience |
Medium |
37601281
|