| 1996 |
A BFA-inhibited guanine nucleotide exchange protein (GEP) for ARF1 and ARF3 was purified from bovine brain cytosol as a ~200 kDa protein containing tryptic peptides 47% identical to yeast Sec7, establishing that ARF3 GDP-to-GTP exchange is catalyzed by a Sec7-domain-containing GEP and is sensitive to brefeldin A. |
Protein purification from bovine brain cytosol (DEAE-Sephacel, hydroxylapatite, Mono Q, Superose 6), SDS/PAGE, silver staining, electroelution/renaturation, in vitro nucleotide exchange activity assay, tryptic peptide sequencing |
Proceedings of the National Academy of Sciences of the United States of America |
High |
8917509
|
| 2000 |
Three residues of human ARF3 (F51, W66, Y81) form a hydrophobic pocket in the GDP-bound state; mutations at these residues increased the rate of GDP dissociation and association but not GTPγS binding, and selectively impaired binding to different ARF effectors in two-hybrid assays, indicating these residues regulate nucleotide exchange kinetics and effector interactions. |
Site-directed mutagenesis of ARF3, in vitro nucleotide dissociation/association assays, yeast two-hybrid effector binding assays |
FEBS letters |
High |
11150519
|
| 2010 |
ARF3 localizes selectively to the trans-Golgi network (TGN) in a manner uniquely dependent on BIG family guanine nucleotide exchange factors; BIGs knockdown redistributes ARF3 but not ARF1 from Golgi membranes, and this TGN association is temperature-sensitive. Mutational analysis identified pairs of residues at opposite ends of ARF3 that separately control membrane recruitment and temperature-dependent release. |
siRNA knockdown of BIG GEFs, fluorescence microscopy, temperature-shift experiments (20°C block), site-directed mutagenesis of ARF3, phylogenetic analysis |
Molecular biology of the cell |
High |
20357002
|
| 2012 |
ARF1 and ARF3 localize to endosomal compartments containing endocytosed transferrin, and simultaneous siRNA depletion of both ARF1 and ARF3 induces tubulation of recycling endosomes (Rab4+, Rab11+, TfR+) and suppresses transferrin recycling to the plasma membrane, without affecting Golgi integrity, early/late endosomes, or retrograde TGN transport. |
siRNA double knockdown of ARF1 and ARF3, EGFP-tagging and fluorescence microscopy, transferrin recycling assay, endocytosis assay, retrograde transport assays (TGN38, CD4-furin, EGF) |
Cell structure and function |
High |
22971977
|
| 2015 |
Class I ARFs (ARF1 and ARF3) localize to the Flemming body during late cytokinesis; double knockdown of ARF1 and ARF3 increases multinucleate cells, and simultaneous triple knockdown of ARF1, ARF3, and ARF6 causes severe cytokinesis defects. EFA6 exchange factor activates both ARF6 and ARF1 in cells. |
siRNA knockdown (double and triple), fluorescence microscopy of Flemming body localization, multinucleate cell quantification, cytokinesis assays |
Journal of biochemistry |
Medium |
26330566
|
| 2020 |
BIG1 (a BFA-inhibited GEF) activates ARF3 in macrophages; BIG1 deficiency inhibits ARF3 activation, reduces PI(4,5)P2 synthesis by impairing PIP5K activation, and prevents TIRAP recruitment to the plasma membrane, thereby suppressing TLR4-MyD88 signaling during LPS stimulation. |
Myeloid-specific BIG1 knockout mouse, siRNA knockdown in bone marrow-derived macrophages and THP-1 cells, ARF3 activation assay, PI(4,5)P2 measurement, TIRAP recruitment assay, cytokine measurement |
Cell death & disease |
Medium |
32415087
|
| 2021 |
De novo missense variants in ARF3 (p.Asp67Val and p.Arg99Leu) cause neurodevelopmental disorder. In vitro assays showed p.Asp67Val causes cytosolic dispersal of ARF3 and dispersed Golgi (loss-of-function), while p.Arg99Leu localizes normally to Golgi but shows increased binding to GGA1 (gain-of-function). In vivo, p.Asp67Val expression was lethal in Drosophila, and p.Arg99Leu caused rough eye phenotype similar to known gain-of-function variant p.Gln71Leu. |
In vitro subcellular localization assays, pull-down assays for GGA1 binding, Drosophila in vivo expression experiments |
Human molecular genetics |
High |
34346499
|
| 2022 |
De novo missense variants in ARF3 affecting the guanine nucleotide binding pocket variably perturb protein stability and GTP/GDP binding, disrupt Golgi morphology and vesicle assembly/trafficking in cell-based assays, and in zebrafish models cause dominant effects on brain size (microcephaly) and body plan formation, impairing neural precursor proliferation and planar cell polarity-dependent cell movements. |
Cell-based assays (Golgi morphology, vesicle trafficking), nucleotide binding assays, zebrafish disease modeling, in vivo analysis of neural precursor proliferation and cell polarity |
Nature communications |
High |
36369169
|
| 2023 |
ARF3 controls the modality of collective cancer cell invasion by associating with and regulating the turnover of N-cadherin; ARF3 loss or gain switches between leader-cell-led chain invasion and collective sheet movement. In vivo, ARF3 levels act as a rheostat for metastasis from intraprostatic tumor transplants. |
Functional genomic screen of 3D prostate cancer cell behavior, siRNA/overexpression, N-cadherin co-association and turnover assays, intraprostatic in vivo tumor transplant model |
The Journal of cell biology |
Medium |
36880595
|
| 2025 |
ARF3 knockdown in mice and cells suppressed influenza A virus (H3N2) replication in vitro and mitigated IAV-induced lung injury in vivo, reducing pro-inflammatory cytokines and attenuating NLRP3 inflammasome activation. |
siRNA knockdown in vitro, mouse IAV pneumonia model, cytokine measurement, NLRP3 inflammasome activation assay |
Virus genes |
Low |
40608252
|