| 1993 |
Vps15 protein kinase forms a hetero-oligomeric membrane-associated complex with the Vps34 PI 3-kinase; Vps15 is responsible for recruiting Vps34 to intracellular membranes, and an intact Vps15 kinase domain is required for activation of Vps34 PI 3-kinase activity. |
Chemical cross-linking, native immunoprecipitation, subcellular fractionation, sucrose density gradients, kinase domain mutagenesis, genetic suppression assay |
The EMBO journal |
High |
8387919
|
| 1995 |
Active Vps15p kinase domain is necessary for physical association with Vps34p and for recruitment of Vps34p to the membrane; catalytically inactive Vps15p mutants cannot associate with Vps34p. Functional Vps15p-Vps34p complex is absolutely required for efficient vacuolar protein delivery, as shown by temperature-conditional alleles causing loss of PI(3)P and missorting of soluble vacuolar proteins. |
Temperature-conditional allele analysis, in vivo PtdIns(3)P measurements, genetic dominant-negative analysis, mutagenesis of kinase/lipid kinase domains |
The Journal of cell biology |
High |
7721937
|
| 1991 |
Alterations of conserved protein kinase residues in Vps15p biologically inactivate the protein, blocking vacuolar sorting of soluble hydrolases (CPY, PrA) but not the membrane protein alkaline phosphatase. C-terminal truncations of Vps15p abolish in vivo phosphorylation of Vps15p and cause a temperature-conditional vacuolar protein sorting defect, suggesting autophosphorylation is required for full activity. |
Site-directed mutagenesis of conserved kinase residues, in vivo 32P-phosphorylation assay, temperature-shift experiments, pulse-chase sorting assay |
The EMBO journal |
High |
1756716
|
| 2009 |
The Vps15 WD-repeat domain forms a seven-bladed beta-propeller resembling a G-protein beta subunit and is sufficient to bind the G protein alpha subunit Gpa1 (preferentially GDP-bound) and Atg14. The kinase and intermediate domains of Vps15 additionally contribute to Gpa1 binding and are necessary for G protein signaling at endosomes. |
X-ray crystal structure of the Vps15 WD domain, binding assays (Gpa1-Vps15 interaction), domain deletion analysis, G protein signaling assays |
Biochemistry |
High |
19445518
|
| 2010 |
A specific PI3K-III sub-complex containing VPS15, VPS34, Beclin 1, UVRAG and BIF-1 (but not ATG14L) regulates endocytic receptor degradation and cytokinesis in mammalian cells. |
siRNA depletion of individual subunits, high-content microscopy-based assays for receptor degradation and cytokinesis, midbody localization of UVRAG and BIF-1 |
Experimental cell research |
Medium |
20643123
|
| 2008 |
Drosophila Vps15 is required for starvation-induced autophagy in fat body and gut; loss of vps15 leads to accumulation of ubiquitin- and Ref(2)P/p62-positive, partially detergent-insoluble protein aggregates, establishing Vps15 as essential for autophagic clearance of protein aggregates in a multicellular organism. |
Drosophila deletion mutant, GFP-Atg8a fluorescence microscopy, electron microscopy, biochemical fractionation, Western blot, immuno-electron microscopy |
Autophagy |
High |
18326940
|
| 2013 |
Vps15 is required for lysosomal function in skeletal muscle; muscle-specific Vps15 knockout mice show accumulation of autophagosomes, p62, LC3, and Lamp2-positive vesicles, and develop autophagic vacuolar myopathy. Importantly, autophagosome formation (LC3-positive) and mTOR activation are maintained in Vps15-deficient cells, indicating Vps15/Vps34 is specifically required downstream (lysosomal fusion/degradation) rather than for autophagosome initiation per se. |
Conditional (muscle-specific) Vps15 knockout mouse, immunofluorescence, electron microscopy, LC3/p62/LAMP2 Western blot, creatine kinase assay, Vps15 rescue by overexpression in Danon disease myoblasts |
EMBO molecular medicine |
High |
23630012
|
| 2016 |
VPS15 interacts with the golgin GM130 at the cis-Golgi, forming a complex devoid of VPS34, and is required for IFT20 release from the Golgi for transport to the primary cilium. A patient missense mutation VPS15-R998Q impairs this Golgi trafficking function without fully disrupting VPS34-dependent activities. VPS15 regulates primary cilium length in human fibroblasts. |
Missense mutation identification in ciliopathy patients, co-immunoprecipitation of VPS15 with GM130, subcellular localization (Golgi), humanized yeast complementation assay, IFT20 localization in patient fibroblasts, zebrafish cilia assay |
Nature communications |
High |
27882921
|
| 2018 |
A hypomorphic Vps15 mutation in mice causes defects in endosomal-lysosomal trafficking and autophagy, resulting in upregulation of Nischarin, which inhibits Pak1 signaling, thereby disrupting neuronal migration and causing hippocampal pyramidal cell layer defects. Complete Vps15 ablation causes accumulation of autophagic substrates, apoptosis, and severe cortical atrophy. |
ENU-induced mouse mutant, conditional knockout, neuronal migration assays, Western blot for Nischarin and Pak1 phosphorylation, epistasis between Vps15, Nischarin, and Pak1 pathways |
Nature neuroscience |
High |
29311744
|
| 2021 |
ULK1/2 kinases phosphorylate VPS15 at six sites (including serine 861 as the major site); mutation of these sites reduces VPS34 activity in vitro and impairs autophagosome formation in cells. VPS15 knockout also reveals ULK-dependent starvation-independent accumulation of ULK substrates and kinase activity-regulated recruitment of autophagy proteins to ubiquitin-positive structures. |
Unbiased phosphoproteomics in Ulk1/2 double-knockout MEFs, in vitro VPS34 kinase assay with VPS15 phosphosite mutants, autophagosome formation assays, VPS15 knockout cell lines |
The EMBO journal |
High |
34121209
|
| 2025 |
Cryo-EM structure of the PI3KC3-C1 complex reveals that the VPS15 pseudokinase domain binds GTP and sequesters its covalently-linked N-terminal myristate in the N-lobe, stabilizing the inactive conformation of VPS34. Upon membrane binding (activation), myristate is liberated, disrupting the inhibitory interaction and enabling VPS34 to catalyze PI3P production on membranes. |
Cryo-electron microscopy of full PI3KC3-C1 complex, structural analysis of pseudokinase-GTP interaction, myristate sequestration/release mechanism, functional validation of VPS34 activation |
Science (New York, N.Y.) |
High |
39913640
|
| 1999 |
Pichia pastoris Vps15 (PpVPS15) is required at an early stage of selective peroxisome autophagy (pexophagy); deletion of PpVPS15 prevents vacuolar uptake of peroxisomes upon glucose or ethanol exposure. |
Gene deletion in Pichia pastoris, measurement of peroxisomal marker enzyme levels, electron microscopy of peroxisome uptake |
Current genetics |
Medium |
10591966
|
| 2014 |
Drosophila Vps15 is required for autophagy induced by multiple stresses (nutrient deprivation, hypoxia, oxidative stress) and for developmentally programmed autophagy in fat body, intestine, and salivary gland. Additionally, Vps15 is necessary for efficient protein secretion from salivary glands, demonstrating a role in secretory vesicle trafficking beyond autophagy. |
Drosophila vps15 genetic mutants, autophagy markers (Atg8, lysotracker), electron microscopy, salivary gland secretion assay |
Cell death and differentiation |
Medium |
25342466
|
| 2019 |
VPS15 knockdown in HUVECs inhibits AngII-induced autophagy and reduces phosphorylation of PDK1 and PKC substrates, while re-expression of Vps15 rescues autophagy; PDK1 and PKC inhibitors phenocopy Vps15 knockdown, placing Vps15 upstream of a PDK1/PKC signaling axis in autophagy regulation. |
shRNA knockdown of Vps15, MDC staining, LC3-II/I Western blot, pharmacological inhibition of PDK1/PKC, rescue by Vps15 re-expression, flow cytometry for apoptosis |
Life sciences |
Low |
31356904
|
| 2025 |
Loss of Vps15 (core PI3K-III subunit) in Drosophila wing imaginal discs selectively impairs an endocytosis-dependent pool of Wingless (Wg) secretion, causing apical accumulation of Wg in secreting cells, while a glypican-mediated Wg pool is unaffected. Evi/Wls cargo receptor undergoes proteasome-dependent degradation rather than accumulation in PI3K-III mutant cells. |
In vivo CRISPR-Cas9 kinome/phosphatome screen, endogenously fluorescently tagged Wg, super-resolution microscopy, ex vivo pharmacological treatments (proteasome inhibitor) |
bioRxivpreprint |
Medium |
bio_10.1101_2025.05.20.655092
|