| 2008 |
ATG14 (mammalian Atg14) is a component of a distinct Beclin 1-Vps34-p150 PI3-kinase complex that is separate from the UVRAG-containing complex; ATG14 and UVRAG bind Beclin 1 in a mutually exclusive manner. The coiled-coil region of ATG14 required for binding Vps34 and Beclin 1 is essential for autophagy. ATG14 localizes to isolation membranes/phagophores during starvation, and its silencing suppresses autophagosome formation. |
Co-immunoprecipitation, subcellular fractionation/localization, siRNA knockdown, computational homology identification |
Molecular biology of the cell |
High |
18843052
|
| 2009 |
ATG14L (Atg14L) and Rubicon are two Beclin 1-binding proteins that form distinct complexes with Beclin 1-hVps34; ATG14L and UVRAG bind Beclin 1 mutually exclusively while Rubicon binds only UVRAG complexes. GFP-ATG14L localizes to the isolation membrane, autophagosome, and ER. Knockout of ATG14L in mouse ES cells causes a defect in autophagosome formation. |
Co-immunoprecipitation, GFP-fusion live imaging, gene knockout in mouse ES cells |
Nature cell biology |
High |
19270696
|
| 2009 |
ATG14L enhances Vps34 lipid kinase activity and upregulates autophagy. Beclin 1 and ATG14L synergistically promote formation of double-membraned organelles associated with Atg5 and Atg12. The Beclin 1-Vps34-p150-UVRAG-ATG14L complex was identified in vivo by mouse genetics and biochemistry. |
In vitro lipid kinase assay, co-immunoprecipitation from mouse tissue, forced expression/dominant-negative studies, electron microscopy |
Nature cell biology |
High |
19270693
|
| 2010 |
ATG14L recruits the autophagy-specific PI3-kinase complex to the ER. ATG14L localizes to the ER and a point mutation causing defective ER localization also abolishes autophagy induction. Knockdown of ATG14L eliminates the DFCP1-positive omegasome. Restoring ER targeting by fusing the DFCP1 ER-targeting motif to the ER-localization-defective ATG14L mutant fully rescues autophagy in ATG14L-knockout ES cells. |
Point mutagenesis, domain-swap rescue, siRNA knockdown, fluorescence imaging of omegasomes, ATG14L KO ES cells |
The Journal of cell biology |
High |
20713597
|
| 2011 |
The C-terminal BATS (Barkor/ATG14(L) autophagosome targeting sequence) domain of ATG14L targets PI3KC3 to autophagosomal membranes by binding to the membrane via an amphipathic alpha helix and preferentially recognizing highly curved membranes enriched in PtdIns(3)P. The BATS domain is required for PI3KC3 recruitment and autophagy stimulation. |
Bioinformatics, site-directed mutagenesis, GFP-fusion imaging, in vitro membrane-binding biochemical assay, membrane curvature sensing assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
21518905
|
| 2012 |
The Beclin 1 coiled-coil domain forms a metastable antiparallel homodimer with 'imperfect' a-d' pairings; ATG14L promotes transition from the metastable Beclin 1 homodimer to a Beclin1-ATG14L heterodimer. Beclin 1 mutants with enhanced self-interaction show altered binding to ATG14L. |
X-ray crystal structure of Beclin 1 CCD, co-immunoprecipitation, mutagenesis |
Nature communications |
High |
22314358
|
| 2012 |
ATG14L binds to the SNARE effector protein Snapin and colocalizes with Snapin to facilitate endosome maturation. ATG14L knockdown delays late-stage endocytic trafficking (retarded receptor degradation kinetics). The Snapin-binding but not Beclin 1-binding activity of ATG14L is required for this endocytic function. |
Co-immunoprecipitation, knockdown, rescue with wild-type vs. binding mutants, surface receptor degradation kinetics assay |
Journal of cell science |
Medium |
22797916
|
| 2013 |
ATG14 controls an autophagy-dependent phosphorylation of Beclin 1 at serines 90 and 93; phosphorylation at these sites is necessary for maximal autophagy. A unique membrane-association domain in Beclin 1 controls autophagosome size and number. |
Mutagenesis of phosphorylation sites, Western blot for phospho-Beclin 1, autophagy flux assays |
Molecular and cellular biology |
Medium |
23878393
|
| 2014 |
NRBF2 directly binds ATG14L through its MIT domain and is a component of the ATG14L-Beclin 1-Vps34-Vps15 complex; NRBF2 enhances ATG14L-linked Vps34 kinase activity and promotes autophagy induction. NRBF2-deficient mice show impaired ATG14L-linked Vps34 activity and focal liver necrosis. |
Co-immunoprecipitation, in vitro kinase assay, NRBF2 knockout mice, domain binding analysis (MIT domain) |
Nature communications |
High |
24849286
|
| 2014 |
Nrbf2 interacts with ATG14L and modulates ATG14L-Vps34/Vps15 interactions to suppress Vps34 activity and reduce PI3P levels, thereby suppressing autophagy. Nrbf2 deficiency increases intracellular PI3P and diminishes ATG14L-Vps34/Vps15 interactions. |
Co-immunoprecipitation, PI3P measurement, siRNA knockdown, isolation membrane marker co-localization |
The Journal of biological chemistry |
Medium |
25086043
|
| 2014 |
Dapper1 (Dpr1) directly interacts with both Beclin 1 and ATG14L and enhances Beclin1-Vps34 interaction and Vps34 kinase activity to promote autophagosome formation. Loss of Dpr1 in the CNS causes motor coordination defects and p62/ubiquitinated protein accumulation. |
Co-immunoprecipitation, pulldown, Vps34 kinase assay, conditional CNS knockout mice, LC3/Atg14L/DFCP1 puncta assay |
Cell research |
Medium |
24980960
|
| 2015 |
ATG14 promotes membrane tethering of protein-free liposomes and enhances hemifusion and full fusion of proteoliposomes reconstituted with t-SNAREs STX17/SNAP29 and v-SNARE VAMP8. ATG14 binds to the SNARE core domain of STX17 through its coiled-coil domain and stabilizes the STX17-SNAP29 binary t-SNARE complex on autophagosomes. ATG14 homo-oligomerization via cysteine repeats is required for membrane tethering, fusion enhancement, and autophagosome-endolysosome fusion in cells; cells expressing homo-oligomerization-defective ATG14 form autophagosomes normally but are blocked in fusion with endolysosomes. |
In vitro reconstituted membrane fusion assay, proteoliposome tethering assay, co-immunoprecipitation, mutagenesis of cysteine repeats, cell biology with autophagosome-lysosome fusion readout |
Nature |
High |
25686604
|
| 2015 |
ATG14 directly binds the STX17-SNAP29 binary complex on autophagosomes and promotes STX17-SNAP29-VAMP8-mediated autophagosome fusion with lysosomes. ATG14 homo-oligomerization is required for SNARE binding and fusion promotion but is dispensable for PI3K stimulation and autophagosome biogenesis. |
Co-immunoprecipitation, biochemical pulldown, cell biology fusion assays, ATG14 homo-oligomerization mutants |
Autophagy |
High |
25945523
|
| 2015 |
ATG14L is ubiquitinated and degraded via the ZBTB16-Cullin3-Roc1 E3 ubiquitin ligase complex. A wide range of GPCR ligands regulate ATG14L levels through ZBTB16, linking GPCR activation to suppression of autophagy. |
Co-immunoprecipitation of ZBTB16-Cullin3-Roc1 complex with ATG14L, ubiquitination assay, pharmacological GPCR manipulation, mouse model of Huntington's disease |
eLife |
Medium |
25821988
|
| 2016 |
ULK1 phosphorylates ATG14 at serine 29 in an mTOR-dependent manner via interaction mediated by ATG13 (which bridges ULK1 to the ATG14-containing PI3K complex). This phosphorylation stimulates ATG14-Vps34 lipid kinase activity and facilitates phagophore and autophagosome formation. ATG14 phosphorylation is decreased in Huntington's disease models due to p62-mediated sequestration of ULK1. |
Phospho-specific antibody generation, in vitro kinase assay, mutagenesis (S29A), co-immunoprecipitation, HD mouse models (Q175), proteasomal inhibition experiments |
Molecular neurodegeneration |
High |
27938392
|
| 2016 |
ATG13 mediates the interaction between ULK1 and the ATG14-containing PIK3C3/VPS34 complex. ULK1 phosphorylates ATG14 in a starvation/MTORC1-inhibition-dependent manner, stimulating PI3K complex kinase activity and autophagosome formation. ATG9A is required to suppress ULK1 activity under nutrient-rich conditions. |
Co-immunoprecipitation, in vitro kinase assay, phospho-ATG14 detection, ATG9A knockdown, ULK1 kinase assays in nutrient-deprivation conditions, in vivo dietary experiments |
Autophagy |
High |
27046250
|
| 2016 |
ATG14's BATS domain also strongly binds PtdIns(4,5)P2. ATG14 specifically interacts with PIPKIγi5, an enzyme generating PtdIns(4,5)P2. PtdIns(4,5)P2 binding to the ATG14-BATS domain regulates ATG14 interaction with VPS34 and Beclin 1 and plays a key role in ATG14 complex assembly and autophagy initiation. Loss of PIPKIγi5 results in loss of ATG14/UVRAG/Beclin 1 and blocks autophagy. |
Co-immunoprecipitation, lipid-binding assay, PIPKIγi5 knockdown/overexpression, autophagy flux measurement |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
27621469
|
| 2016 |
PAQR3 functions as a scaffold protein that preferentially facilitates the formation of ATG14L-linked (but not UVRAG-linked) VPS34 complex. Upon glucose starvation, AMPK phosphorylates PAQR3 at threonine 32, switching on PI3P production via the ATG14-associated PI3K complex. Loss of PAQR3 reduces exercise-induced autophagy and ATG14L-associated VPS34 complex integrity. |
Co-immunoprecipitation, in vitro AMPK kinase assay, site-directed mutagenesis (T32), PI3P measurement, Paqr3 knockout mice |
The EMBO journal |
High |
26834238
|
| 2016 |
SLC35D3, an ER-associated transmembrane protein expressed in midbrain dopaminergic neurons, enhances the formation of the BECN1-ATG14-PIK3C3 complex to induce autophagy. Loss of SLC35D3 (ros mutant mice) reduces dopaminergic neurons and impairs autophagy. |
Co-immunoprecipitation, ros mutant mouse analysis, dopaminergic neuron counting, autophagy flux measurement |
Autophagy |
Medium |
27171858
|
| 2018 |
ULK1 phosphorylates Beclin 1 at Ser30 specifically in association with ATG14 (but not with UVRAG). This phosphorylation activates the ATG14-containing PIK3C3/VPS34 kinase complex and stimulates autophagosome formation in response to amino acid starvation, hypoxia, and MTORC1 inhibition. The S30A Beclin 1 mutation does not affect ULK1-mediated phosphorylation of Beclin 1 Ser15 or ATG14 Ser29. |
In vitro kinase assay, phospho-specific antibody, Ser30Ala mutagenesis, co-immunoprecipitation, autophagy flux and phagophore formation assays |
Autophagy |
High |
29313410
|
| 2018 |
ULK1 O-GlcNAcylation at threonine 754 by OGT upon glucose starvation is required for binding and phosphorylation of ATG14L, enabling VPS34 activation, PI3P production, and autophagy initiation. |
O-GlcNAcylation site mapping, mutagenesis (T754), co-immunoprecipitation of ULK1-ATG14L, VPS34 kinase assay, PI3P measurement |
Cell reports |
Medium |
30517873
|
| 2016 |
The BECN1 CCD forms an antiparallel homodimer (1.46 Å crystal structure); the ATG14 CCD is largely disordered alone but becomes more helical upon forming a parallel BECN1:ATG14 CCD heterodimer. The heterodimer is more curved than other BECN1-containing dimers. BECN1 homodimer interface residues also mediate ATG14 heterodimerization; mutations at these residues abrogate starvation-induced but not basal autophagy. |
X-ray crystallography (1.46 Å), SAXS, circular dichroism, co-immunoprecipitation, autophagy flux assays with mutants |
Biochemistry |
High |
27383850
|
| 2012 |
FoxO transcription factors and core clock machinery (Clock/Bmal1) regulate Atg14 gene expression. FoxO-binding cis-elements and E-box elements are present in the Atg14 proximal promoter. Hepatic Atg14 knockdown elevates triglycerides in liver and serum; overexpression of Atg14 improves hypertriglyceridemia in high-fat diet mice and FoxO1/3/4 liver-specific KO mice. |
Luciferase reporter assay, chromatin immunoprecipitation, in vivo gene knockdown/overexpression in mouse liver, triglyceride measurement |
The Journal of biological chemistry |
Medium |
22992773
|
| 2017 |
BECN2 CCD forms a metastable antiparallel homodimer; the BECN2 CCD forms a parallel heterodimer with the ATG14 CCD that is stabilized by conserved polar interactions. BECN2 binds ATG14 more tightly than BECN1. These structural differences dictate competitive ATG14 recruitment. |
X-ray crystallography, co-immunoprecipitation, mutagenesis, thermal stability assays |
Protein science |
High |
28218432
|
| 2019 |
ATG14 interacts with Ulk1 and LC3. ATG14 contains a phosphatidylethanolamine (PE)-binding region; Ulk1 addition decreases ATG14-PE interaction, which is concomitant with increased LC3 lipidation, suggesting that Ulk1 sorts PE to LC3 during ATG14-mediated lipophagy. |
Co-immunoprecipitation, PE-binding assay, confocal microscopy, knockdown of Ulk1 |
Free radical biology & medicine |
Low |
28069524
|
| 2019 |
Conditional deletion of ATG14 in the intestinal epithelium causes spontaneous villus loss and increased enterocyte apoptosis, rescued by TNF-blocking antibody or genetic deletion of Tnfr1. This places ATG14-dependent autophagy upstream of TNF-triggered apoptosis in intestinal homeostasis. |
Conditional KO (villin-Cre), histology, apoptosis markers, genetic epistasis with Tnfr1 KO, TNF-blocking antibody treatment |
Autophagy |
High |
30894050
|
| 2020 |
GLIPR2 directly binds purified PtdIns3K-C1 (containing ATG14) and inhibits its in vitro lipid kinase activity. CRISPR-mediated GLIPR2 depletion increases autophagic flux and PI3P generation, identifying GLIPR2 as a negative regulator of the ATG14-containing PI3K complex. |
In vitro PI3K lipid kinase assay with purified PtdIns3K-C1, CRISPR-Cas9 KO, PI3P measurement, autophagy flux assay, WIPI2 recruitment |
Autophagy |
High |
33222586
|
| 2020 |
Disruption of the Beclin 1-ATG14L protein-protein interaction by a small molecule selectively disrupts VPS34 Complex I formation and inhibits autophagy without affecting the UVRAG-Beclin 1 interaction (Complex II) or vesicle trafficking. |
NanoBRET cellular assay, high-throughput screen, co-immunoprecipitation of VPS34 complex components, autophagy flux readout |
Journal of the American Chemical Society |
Medium |
32320221
|
| 2021 |
ATG14 is essential for restricting Mycobacterium tuberculosis replication in human iPSC-derived macrophages; ATG14 deletion increases replication of both wild-type Mtb and a mutant (ΔesxBA) that cannot trigger canonical autophagy. ATG14 regulates fusion of phagosomes containing Mtb with lysosomes. |
CRISPR-Cas9 deletion in human iPSDM, single-cell high-content imaging, Mtb replication quantification, phagosome-lysosome fusion assay |
Nature microbiology |
High |
36959508
|
| 2021 |
Muscle-specific conditional KO of ATG14 leads to hypertrophic cardiomyopathy with abnormal accumulation of autophagic cargoes in the heart and early mortality, and autophagic vacuolar myopathy with ubiquitin+/SQSTM1+ deposits in skeletal muscle. |
Conditional KO (Ckm-Cre), histology, autophagy cargo accumulation assays, cardiac function measurement |
Autophagy |
High |
33794726
|
| 2022 |
SETD2 promotes expression of the long ATG14 isoform (ATG14L) that contains N-terminal cysteine repeats essential for efficient autophagosome-lysosome fusion; the short isoform ATG14S lacks this domain. SETD2 loss decreases autophagic flux and impairs clearance of mutant HTT. |
SETD2 loss-of-function, isoform-specific expression analysis, autophagy flux assays, mHTT clearance assay |
Cell death & disease |
Medium |
36371383
|
| 2022 |
Miga (Drosophila ortholog), a mitochondrial outer-membrane protein regulating ER-mitochondria contact sites, binds ATG14 and Uvrag. Miga-mediated stabilization of Syx17 (STX17 ortholog) is dependent on Atg14. Miga-regulated ERMCSs are critical for PI3P formation but not for Syx17 stabilization. |
Co-immunoprecipitation, genetic epistasis in Drosophila, PI3P measurement, Syx17 protein stability assay |
Cell reports |
Medium |
36323251
|
| 2023 |
MARCH7 (MARCHF7), an E3 ubiquitin ligase, ubiquitinates ATG14 with K6-, K11-, and K63-linked mixed polyubiquitin chains, triggering ATG14 aggregation and reducing its solubility. Ubiquitinated ATG14 has fewer interactions with STX17, inhibiting autophagy flux. MARCH7 depletion decreases aggresome-like induced structures (ALISs). |
Co-immunoprecipitation, ubiquitination assay with linkage-specific analysis, solubility fractionation, STX17 co-IP with ubiquitination mutants, MARCH7 KO cells |
Cell reports |
High |
37632749
|
| 2023 |
RUNDC1 is a novel ATG14-interacting protein that negatively modulates autophagy by blocking autophagosome-lysosome fusion. RUNDC1 stimulates ATG14 homo-oligomerization to prevent VAMP8 from binding STX17-SNAP29, clasping the ATG14-STX17-SNAP29 complex. Phosphorylation of RUNDC1 Ser379 is crucial for this inhibitory function. |
Co-immunoprecipitation, gain/loss-of-function in human cells and zebrafish, SNARE complex assembly assay, phospho-site mutagenesis |
Cell death and differentiation |
High |
37684417
|
| 2023 |
Streptococcus pneumoniae CbpC interacts with the coiled-coil domain of ATG14 (Y83 of CbpC dp3 domain is critical), activating autophagy; simultaneously CbpC deploys ATG14 for autophagic degradation via ATG14-CbpC-p62/SQSTM1 axis, reducing ATG14-STX17 complex formation and suppressing antibacterial autophagy. |
Co-immunoprecipitation, domain mutagenesis, bacterial survival assay, ATG14-STX17 complex analysis |
EMBO reports |
Medium |
32239622
|
| 2024 |
ATG14 targets lipid droplets (LDs) via its BATS domain and acts as an autophagic receptor for LD degradation (lipophagy) by interacting with ATG8 family proteins. STX18 (syntaxin 18) binds ATG14, disrupting ATG14-ATG8 interactions and PI3KC3-C1 complex formation. Coronavirus M protein binds STX18 to subvert the STX18-ATG14 interaction, inducing lipophagy and degrading the antiviral protein Viperin to facilitate virus production. |
Co-immunoprecipitation, domain mutagenesis (BATS domain), siRNA knockdown, fluorescence imaging of LDs, in vitro binding assays, viral replication assay |
Nature communications |
High |
38245527
|
| 2024 |
ATG14's BATS domain is responsible for ATG14 localization to lipid droplets. ATG14 interacts with ATGL (adipose triglyceride lipase) and CGI-58 and enhances the ATGL-CGI-58 interaction. ATG14 deficiency decreases triglyceride hydrolysis in vitro, indicating ATG14 directly promotes lipolysis at lipid droplets. |
Fluorescence imaging, BATS domain mutagenesis, co-immunoprecipitation of ATGL and CGI-58, in vitro lipolysis assay, ATG14 knockdown in hepatocytes and mouse livers |
Metabolism: clinical and experimental |
High |
37741434
|
| 2022 |
PI4K2A generates a pool of PI4P that facilitates recruitment of ATG14 to mature autophagosomes. PI4K2A binds to ATG14. Impaired ATG14 targeting to autophagosomes in PI4K2A-depleted cells is rescued by introduction of PI4P but not PI(4,5)P2, indicating PI4P and PI(4,5)P2 have independent functions in late-stage autophagy. |
PI4K2A depletion, PI4P/PI(4,5)P2 rescue experiments, co-immunoprecipitation, ATG14 localization imaging |
Biochemistry |
Medium |
35380781
|
| 2024 |
GULP1 interacts with ATG14 and potentiates the stimulatory effect of ATG14 on PI3KC3-C1 activity. GULP1 facilitates targeting of ATG14 to the ER. GULP1 promotes APP entry into autophagic vacuoles and enhances APP processing; a GULP1 mutant disrupting the GULP1-ATG14 interaction attenuates these effects. |
Co-immunoprecipitation, PI3KC3-C1 kinase assay, ATG14-ER localization imaging, APP processing assay, GULP1 mutant rescue |
Cellular and molecular life sciences |
Medium |
39080084
|
| 2025 |
USP1 deubiquitinates ATG14, stabilizing it by reducing bound ubiquitin levels and preventing proteasome-dependent degradation. USP1 depletion or inhibition reduces autophagy initiation and flux. USP1 is the first identified deubiquitinase for ATG14. |
Co-immunoprecipitation, ubiquitination assay, USP1 overexpression/knockdown, proteasome inhibition rescue, xenograft tumor model |
The Journal of biological chemistry |
Medium |
39814232
|
| 2025 |
Conditional depletion of ATG14 in the mouse oviduct causes severe structural abnormalities, abnormal embryo retention, and impaired embryo transport. Mechanistically, loss of ATG14 triggers unscheduled pyroptosis via mitochondrial integrity disruption. Pharmacological pyroptosis activation phenocopies the ATG14-KO embryo transport defect. |
Conditional KO (oviduct-specific Cre), histology, embryo transport assay, mitochondrial integrity assay, pharmacological pyroptosis induction |
eLife |
Medium |
40100261
|