| 1997 |
VPS29 (yeast) is required for endosome-to-Golgi retrieval of the vacuolar sorting receptor Vps10p; in vps29 mutants, Vps10p mislocalizes from the Golgi to the vacuolar membrane, indicating VPS29 functions in retrograde recycling of cargo receptors from the prevacuolar endosome back to the Golgi. |
Genetic loss-of-function (yeast vps29 mutants), subcellular fractionation, CPY secretion assay |
The Journal of cell biology |
High |
9105038
|
| 2005 |
Crystal structure of mammalian VPS29 reveals a phosphoesterase fold with structural similarity to divalent metal-containing phosphoesterases; VPS29 has no detectable phosphoesterase activity in vitro. VPS29 and VPS26 bind independently to VPS35 to form a high-affinity heterotrimeric subcomplex. A conserved hydrophobic surface on VPS29 (distinct from the VPS35-binding site) mediates assembly with sorting nexins in yeast; mutations at either binding site cause defects in retromer-dependent membrane trafficking. |
X-ray crystallography, in vitro phosphatase assay, mutagenesis, binding/assembly assays, in vivo trafficking assay |
Nature structural & molecular biology |
High |
15965486
|
| 2006 |
Recombinant human VPS29 displays in vitro phosphatase activity toward a serine-phosphorylated peptide from the acidic-cluster dileucine motif of the CI-M6PR cytoplasmic tail; activity requires co-presence of VPS26 and VPS35, is abolished by alanine substitution of active-site metal-coordinating residues, and depends on zinc (VPS29 binds Zn²⁺ as shown by ICP-MS; activity is abrogated by zinc chelators and restored by ZnCl₂). |
In vitro phosphatase assay with recombinant proteins, active-site mutagenesis, ICP-MS metal analysis, metal chelation experiments |
The Biochemical journal |
Medium |
16737443
|
| 2011 |
VPS29 is not a functional metalloenzyme: despite coordinating Mn²⁺ and Zn²⁺ in the putative active site, affinity for metals is low and no phosphatase activity is detected toward a putative peptide substrate. NMR/ITC and RDC measurements show VPS29 is a rigid scaffold with metal-independent conformation. NMR chemical shift mapping demonstrates VPS29 associates with SNX1 via a conserved hydrophobic surface, but with low affinity requiring additional interactions in vivo. |
X-ray crystallography, NMR spectroscopy (RDC, chemical shift mapping), ITC, in vitro phosphatase assay |
PloS one |
High |
21629666
|
| 2010 |
VPS29 deficiency leads to degradation of VPS35 (and vice versa), but VPS26 deficiency does not affect VPS29 or VPS35 levels. The VPS29-VPS35 sub-complex is more stable than the VPS26-VPS35 sub-complex in vitro, and the VPS26-VPS35 sub-complex is more susceptible to ubiquitin-proteasome-mediated degradation, indicating VPS29 stabilizes VPS35 as a biologically stable intermediate during retromer assembly. |
siRNA knockdown, in vitro complex formation, ubiquitin-proteasome inhibitor treatment, immunoblotting |
FEBS letters |
Medium |
25937119
|
| 2017 |
The Legionella effector RidL binds the VPS29 retromer subunit through a protruding β-hairpin containing Ile170; this interaction displaces the Rab7 GAP TBC1D5 from VPS29 and from Legionella-containing vacuoles, blocking retrograde vesicle trafficking and promoting intracellular bacterial replication. Deletion of the β-hairpin or substitution of Ile170 (RidL) or Leu152 (VPS29) abolishes the interaction in cells and in vitro. |
Crystal structure of RidL-VPS29 complex, mutagenesis, co-immunoprecipitation in eukaryotic cells, in vitro binding assays, intracellular replication assay |
Nature communications |
High |
29146912
|
| 2020 |
VARP contains a 12-residue, four-cysteine Zn-fingernail microdomain that binds the same hydrophobic site on VPS29 as TBC1D5; VARP and TBC1D5 compete for VPS29 binding in vivo. Mutations abolishing VPS29:VARP binding inhibit trafficking from endosomes to the cell surface. In assembled retromer arches, the geometry of VPS29 subunits favors VARP binding to two VPS29 subunits simultaneously. |
NMR spectroscopy, X-ray crystallography, mutagenesis, in vivo competition assays, endosomal trafficking assays |
Nature communications |
High |
33024112
|
| 2020 |
In Drosophila, Vps29 is required for correct localization of retromer (Vps35/Vps26) in neurons: in Vps29 mutants, Vps35 and Vps26 remain associated but retromer mislocalizes from neuropil to soma together with Rab7. Vps29 mutant phenotypes (impaired synaptic transmission, locomotion defects, endolysosomal dysfunction) are suppressed by reducing Rab7 or overexpressing the Rab7 GAP TBC1D5, placing VPS29 upstream of Rab7 regulation in the retromer pathway. |
Drosophila genetics (loss-of-function, epistasis), immunofluorescence/localization, electrophysiology, behavioral assays, electron microscopy |
eLife |
High |
32286230
|
| 2022 |
VPS29 deficiency in human lung cells causes changes in endosome morphology and acidity and attenuates endosomal protease activity, trapping incoming coronavirus (HCoV-OC43, SARS-CoV-2) particles in endosomes and reducing infection. VPS29 loss had no effect on influenza A endosomal entry and enhanced influenza A infection, demonstrating opposing roles of VPS29-dependent endosome regulation on different viruses. |
Genome-wide CRISPR loss-of-function screen, genetic validation (VPS29 KO), viral infection assays, endosome morphology and acidity measurements, endosomal protease activity assay |
mBio |
Medium |
35229640
|
| 2024 |
VPS29 directly binds a peptide fragment of the WASH complex subunit FAM21 via a conserved hydrophobic pocket on VPS29; the crystal structure shows FAM21 adopts a sharp bend to insert into this pocket using the same binding mode as other VPS29 effectors (e.g., RidL, TBC1D5, VARP). This interaction is distinct from VPS35:FAM21 contacts. The Parkinson's disease-linked VPS35 D620N mutation does not significantly impair direct FAM21 association in vitro. |
Crystal structure of VPS29:FAM21 peptide complex, in vitro binding assays, mutagenesis |
Protein science : a publication of the Protein Society |
High |
38607248
|
| 2025 |
A third VPS29 isoform (VPS29C) harbors an extended N-terminal sequence that constitutes an autoinhibitory domain blocking the hydrophobic groove required for effector recruitment to Retromer and for association with Retriever and Commander complexes. VPS29C is therefore uniquely able to uncouple Retromer-dependent cargo sorting from broader VPS29A/B roles in accessory protein recruitment. |
AlphaFold structural modeling, in vitro complex reconstitution, mass spectrometry, molecular cell biology |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
40587794
|
| 2013 |
In C. elegans, loss of vps-29 (cargo-selective retromer subunit) disrupts Wls recycling from endosomes to TGN, impairing Wnt secretion; this defect can be rescued by blocking late endosomal maturation, which accumulates Wls in late endosomes where it can be retrieved via an SNX-BAR-dependent retromer pathway, demonstrating that the VPS29-containing cargo-selective subcomplex preferentially acts at an early endosomal station distinct from the SNX-BAR pathway. |
C. elegans genetics (vps-29 loss-of-function), Wnt secretion assay, pharmacological inhibition of endosomal maturation, fluorescence microscopy |
Cellular signalling |
Medium |
24056045
|
| 2024 |
VARP directly interacts with SNX27 PDZ domain via its N-terminus, and VARP is required to reconstitute an endosomal supercomplex containing SNX27, ESCPE-1, and Retromer (VPS26/VPS35/VPS29) on PI(3)P-enriched membranes in a fully reconstituted biochemical system; specific point mutations in VARP abolish the VARP:SNX27 interaction in vitro. |
Biochemical reconstitution with purified mammalian proteins, AlphaFold2 Multimer modeling, liposome tubulation assay, in vitro binding assays, mutagenesis |
bioRxivpreprint |
Medium |
|
| 2024 |
ATG5 associates with retromer core components VPS26, VPS29, and VPS35; knockout of ATG5 or other genes essential for membrane atg8ylation impairs retromer-dependent sorting of GLUT1 to the plasma membrane, independently of canonical autophagy, revealing a noncanonical role for membrane atg8ylation in retromer assembly and function. |
Co-immunoprecipitation, ATG5/atg8ylation gene knockouts, GLUT1 trafficking assay |
bioRxivpreprint |
Low |
|