| 2008 |
Human ATG14 (mammalian homologue of yeast Atg14) forms a distinct class III PI3-kinase complex with Beclin 1, Vps34, and p150 that is mutually exclusive with UVRAG-containing complexes. ATG14 localizes to autophagic isolation membranes and its coiled-coil region required for binding Vps34 and Beclin 1 is essential for autophagosome formation; silencing ATG14 in HeLa cells suppresses autophagosome formation. |
Co-immunoprecipitation, subcellular fractionation/localization by fluorescence microscopy, siRNA knockdown with autophagosome formation readout, computational homology identification |
Molecular biology of the cell |
High |
18843052
|
| 2009 |
ATG14L and UVRAG bind to Beclin 1 in a mutually exclusive manner, defining at least three distinct Beclin 1 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-tagged protein localization by fluorescence microscopy, ATG14L knockout in mouse ES cells with autophagosome formation assay |
Nature cell biology |
High |
19270696
|
| 2009 |
ATG14L (Atg14L) enhances Vps34 lipid kinase activity and upregulates autophagy, whereas Rubicon reduces Vps34 activity. ATG14L and Beclin 1 synergistically promote formation of double-membraned organelles associated with Atg5 and Atg12. |
In vivo biochemistry (large Beclin 1 complex characterization), PI3-kinase lipid kinase assay, immunofluorescence for double-membrane organelle formation, mouse genetics |
Nature cell biology |
High |
19270693
|
| 2010 |
ATG14L recruits the class III PI3-kinase complex to the ER, where it generates PI3P to establish the ER as a platform for autophagosome formation. Knockdown of ATG14L eliminates DFCP1-positive omegasomes, and a point mutation causing defective ER localization also ablates autophagy induction; adding an ER-targeting motif to this mutant fully rescues autophagic defects in ATG14L KO ES cells. |
siRNA knockdown, point mutagenesis rescue experiments, ER-targeting domain swap, fluorescence microscopy of DFCP1/omegasome, ATG14L KO ES cell complementation |
The Journal of cell biology |
High |
20713597
|
| 2011 |
The C-terminal BATS (Barkor/ATG14(L) autophagosome targeting sequence) domain of ATG14L is required for autophagosome targeting. BATS contains an amphipathic alpha helix that binds autophagosome membranes enriched in PtdIns(3)P and PtdIns(4,5)P2 and preferentially senses highly curved membranes; deletion of BATS abolishes PI3KC3 recruitment and autophagy stimulation. |
Bioinformatics and mutagenesis of BATS domain, fluorescence microscopy co-localization with Atg16/LC3/DFCP1, in vitro membrane-binding biochemical assay, liposome 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 rendered unstable by 'imperfect' a-d' pairings; ATG14L (and UVRAG) promote transition of this metastable homodimer to stable Beclin1-ATG14L heterodimer. Beclin 1 mutants with enhanced self-interaction show altered binding to ATG14L. |
X-ray crystallography of Beclin 1 CCD, co-immunoprecipitation of mutants, biochemical homodimer/heterodimer formation assays |
Nature communications |
High |
22314358
|
| 2012 |
ATG14L binds to the fusogenic SNARE effector protein Snapin and co-localizes with it to facilitate endosome maturation. The Snapin-binding activity of ATG14L is distinct from its Beclin 1-binding activity; a Snapin-binding mutant of ATG14L fails to rescue the endocytic trafficking delay caused by ATG14L knockdown, while a Beclin 1-binding mutant does rescue it. |
Co-immunoprecipitation, fluorescence co-localization, siRNA knockdown, domain mutant rescue assays measuring receptor degradation kinetics |
Journal of cell science |
Medium |
22797916
|
| 2012 |
ATG14 is critical for 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. |
Phosphorylation site mapping, site-directed mutagenesis of Beclin 1 S90/S93, autophagy flux assays, yeast genetic analysis of Atg6 domain |
Molecular and cellular biology |
Medium |
23878393
|
| 2014 |
NRBF2 directly binds ATG14L through its MIT domain and is a component of the specific ATG14L-Beclin1-Vps34-Vps15 complex. NRBF2 binding to ATG14L enhances ATG14L-linked Vps34 kinase activity and autophagy induction; NRBF2-deficient mice show impaired ATG14L-linked Vps34 activity and autophagy with focal liver necrosis. |
Co-immunoprecipitation, MIT domain binding assay, Vps34 lipid kinase assay, NRBF2 KO mice with liver phenotype analysis |
Nature communications |
High |
24849286
|
| 2014 |
NRBF2 (Nrbf2) interacts and co-localizes with ATG14L within the ATG14L-containing Beclin1-Vps34 complex. Nrbf2 deficiency leads to increased PI3P levels and diminished ATG14L-Vps34/Vps15 interactions, suggesting Nrbf2-mediated ATG14L-Vps34/Vps15 interactions inhibit Vps34 activity and suppress autophagosome biogenesis. |
Co-immunoprecipitation, siRNA knockdown of Nrbf2, PI3P measurement, fluorescence co-localization with isolation membrane markers |
The Journal of biological chemistry |
Medium |
25086043
|
| 2015 |
ATG14 directly binds the SNARE core domain of STX17 (syntaxin 17) through its coiled-coil domain, stabilizing the STX17-SNAP29 binary t-SNARE complex on autophagosomes. ATG14 homo-oligomerization via cysteine repeats is required for membrane tethering and for enhancing hemifusion and full fusion of proteoliposomes reconstituted with STX17, SNAP29, and VAMP8. ATG14 homo-oligomerization-defective cells form autophagosomes normally but block fusion with endolysosomes. |
In vitro reconstituted membrane tethering and fusion assays with protein-free liposomes and proteoliposomes, biochemical SNARE binding assay, homo-oligomerization mutant cell lines, autophagic flux assays |
Nature |
High |
25686604
|
| 2015 |
ATG14 homo-oligomerization is required for binding the STX17-SNAP29 binary SNARE complex on autophagosomes and for promoting autophagosome-lysosome fusion, but is dispensable for PI3K stimulation and autophagosome biogenesis. This was confirmed using biochemical, cell biology, and genetic approaches showing ATG14 homo-oligomerization mutants lose SNARE binding and fusion-promoting activity while retaining PI3K activity. |
Biochemical SNARE complex binding assay, cell biology with homo-oligomerization mutants, genetic complementation, autophagosome-lysosome fusion assay |
Autophagy |
High |
25945523
|
| 2015 |
ATG14L is ubiquitinated and degraded by the ZBTB16-Cullin3-Roc1 E3 ubiquitin ligase complex. A wide range of G-protein-coupled receptor (GPCR) ligands regulate ATG14L protein levels through ZBTB16, providing a mechanism by which GPCR activation suppresses autophagy. |
Co-immunoprecipitation of ATG14L with ZBTB16-Cullin3-Roc1 complex, ubiquitination assay, GPCR ligand treatment with ATG14L protein level measurement, pharmacological autophagy modulation in Huntington's disease mouse model |
eLife |
Medium |
25821988
|
| 2016 |
ULK1 phosphorylates ATG14 at serine 29 in an mTOR-dependent manner, and this phosphorylation critically regulates ATG14-Vps34 lipid kinase activity to control autophagy level. ATG14 phosphorylation is decreased in Q175 Huntington's disease mice partly through p62-induced sequestration of ULK1, and phospho-mimetic ATG14 facilitates clearance of polyQ mutant protein. |
Anti-phospho-ATG14 (Ser29) antibody generation, ULK1 kinase assay, Vps34 lipid kinase activity assay, HD genetic mouse model analysis, proteasomal inhibition studies |
Molecular neurodegeneration |
High |
27938392
|
| 2016 |
The ULK1 complex phosphorylates ATG14 at serine 29 via ATG13-mediated interaction of ULK1 with the ATG14-containing PIK3C3/VPS34 complex. ATG14 phosphorylation stimulates class III PI3K complex kinase activity and facilitates phagophore and autophagosome formation. ULK1-mediated ATG14 phosphorylation requires BECN1 but not PE-conjugation machinery or PIK3C3 kinase activity. ATG9A is required to suppress ULK1 activity under nutrient-enriched conditions. |
Co-immunoprecipitation (ATG13-ULK1-ATG14 interaction), ULK1 kinase assay for ATG14 Ser29 phosphorylation, PI3K activity assay, starvation/MTORC1 inhibition conditions, ATG9A epistasis experiment |
Autophagy |
High |
27046250
|
| 2016 |
PAQR3 functions as a scaffold protein that preferentially facilitates formation of the ATG14L-linked Vps34 complex (not UVRAG-linked), enhancing PI3P generation. AMPK phosphorylates PAQR3 at threonine 32, which is required for activation of ATG14L-associated PI3K upon glucose starvation. |
Co-immunoprecipitation of PAQR3 with ATG14L-Beclin1-Vps34 complex, PI3P generation assay, AMPK phosphorylation site mutagenesis (T32A), PAQR3 deletion mice with exercise-induced autophagy assay |
The EMBO journal |
High |
26834238
|
| 2016 |
ATG14 specifically interacts with type Iγ PIP kinase isoform 5 (PIPKIγi5), an enzyme generating PtdIns(4,5)P2. The BATS domain of ATG14 binds PtdIns(4,5)P2, regulating ATG14 interaction with VPS34 and Beclin 1 and ATG14 complex assembly. Loss of PIPKIγi5 results in loss of ATG14, UVRAG, and Beclin 1 and autophagy block. |
Co-immunoprecipitation of ATG14 with PIPKIγi5, BATS domain lipid-binding assay for PtdIns(4,5)P2, PIPKIγi5 knockdown with ATG14/Beclin1 protein level and autophagy assays |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
27621469
|
| 2016 |
ATG14 crystal structure (BECN1:ATG14 CCD heterodimer) determined by SAXS shows the heterodimer is more curved than other BECN1-containing dimers. ATG14 CCD is disordered alone but becomes more helical upon heterodimerization with BECN1 CCD. Point mutations at the BECN1:ATG14 interface abrogate starvation-induced but not basal autophagy. |
X-ray crystallography of BECN1 CCD homodimer, SAXS of BECN1:ATG14 CCD heterodimer, circular dichroism, co-immunoprecipitation of interface mutants, starvation-induced autophagy assay |
Biochemistry |
High |
27383850
|
| 2017 |
BECN2 CCD forms an antiparallel homodimer with weaker self-association than BECN1 but binds ATG14 CCD more tightly than BECN1 does. Crystal structure shows BECN2:ATG14 CCD forms a parallel, curved heterodimer stabilized by conserved polar interactions; mutation of nonideal BECN2 interface residues improves homodimerization but weakens ATG14 binding. |
X-ray crystallography of BECN2 CCD homodimer and BECN2:ATG14 CCD heterodimer, site-directed mutagenesis, thermal stability assay, binding affinity measurements |
Protein science |
High |
28218432
|
| 2018 |
ULK1 phosphorylates BECN1 at Ser30 specifically in association with ATG14-containing (not UVRAG-containing) PIK3C3 complex. This phosphorylation is induced by amino acid starvation, hypoxia, and MTORC1 inhibition, and requires ATG13 and RB1CC1. Blocking Ser30 phosphorylation (S30A mutant) suppresses ATG14-containing PIK3C3 kinase activity and reduces autophagy flux and autophagosome formation without affecting ATG14 Ser29 phosphorylation. |
In vitro ULK1 kinase assay for BECN1 Ser30, site-directed mutagenesis (S30A), co-immunoprecipitation, PIK3C3 lipid kinase activity assay, autophagy flux measurement |
Autophagy |
High |
29313410
|
| 2018 |
ULK1 is O-GlcNAcylated at threonine 754 by OGT upon glucose starvation (after mTOR-dependent Ser757 dephosphorylation by PP1 and AMPK-mediated phosphorylation). ULK1 O-GlcNAcylation is required for binding and phosphorylation of ATG14L, enabling VPS34 lipid kinase activation, PI3P production, and phagophore formation. |
O-GlcNAc modification identification (mass spectrometry), OGT/PP1 inhibitor studies, co-immunoprecipitation of ULK1-ATG14L, VPS34 lipid kinase assay, phagophore formation assay |
Cell reports |
Medium |
30517873
|
| 2019 |
Intestinal epithelium-specific ATG14 knockout mice develop spontaneous villus loss and epithelial cell death (apoptosis) in the small intestine. ATG14-deficient intestinal epithelial cells are sensitive to TNF-triggered apoptosis; TNF-blocking antibody and genetic deletion of TNFR1 rescue the phenotype. Similarly, FIP200 intestinal-specific deletion causes the same villus atrophy phenotype. |
Conditional KO (Atg14-Cre/VillinCre) mice, caspase cleavage markers, TNF-blocking antibody rescue, TNFR1 genetic deletion epistasis |
Autophagy |
High |
30894050
|
| 2020 |
GLIPR2 (a Golgi-associated protein) binds to the BECN1-ATG14-containing PtdIns3K-C1 complex and directly inhibits its in vitro lipid kinase activity. CRISPR-Cas9 depletion of GLIPR2 increases autophagic flux and PI3P generation in cells and in mice. |
In vitro lipid kinase assay with purified PtdIns3K-C1 plus recombinant GLIPR2, CRISPR-Cas9 KO in cells and mice, PI3P measurement, autophagic flux assay |
Autophagy |
High |
33222586
|
| 2021 |
ATG14 muscle-specific conditional KO (atg14-cKO) mice develop hypertrophic cardiomyopathy with abnormal accumulation of autophagic cargoes in the heart and early mortality; skeletal muscles show autophagic vacuolar myopathy with ubiquitin+/SQSTM1+ deposits. Unlike rb1cc1-cKO mice, atg14-cKO mice do not show TARDBP/TDP-43+ pathology, indicating distinct roles for ATG14 vs. RB1CC1 in muscle autophagy. |
Muscle-specific conditional KO mice (Ckm-Cre), histopathology, immunohistochemistry for autophagy cargo markers, cardiac function assessment |
Autophagy |
High |
33794726
|
| 2022 |
SETD2 promotes expression of the long ATG14 isoform (ATG14L) containing the N-terminal cysteine repeats domain required for efficient autophagosome-lysosome fusion. Loss of SETD2 decreases ATG14L expression and autophagic flux, and impairs degradation of aggregation-prone mutant HTT. |
SETD2 loss-of-function, isoform expression analysis, autophagic flux measurement, mutant HTT clearance assay |
Cell death & disease |
Medium |
36371383
|
| 2022 |
PI4K2A generates a pool of PI4P on mature autophagosomes that facilitates ATG14 recruitment. PI4K2A binds ATG14, suggesting in situ PI4P synthesis near ATG14. Impaired ATG14 targeting to autophagosomes in PI4K2A-depleted cells is rescued by exogenous PI4P but not PI(4,5)P2. |
PI4K2A knockdown with ATG14 localization assay by fluorescence microscopy, PI4K2A-ATG14 co-immunoprecipitation, PI4P and PI(4,5)P2 rescue experiments |
Biochemistry |
Medium |
35380781
|
| 2022 |
Drosophila Miga (ER-mitochondrial contact protein) binds Atg14 and Uvrag; Miga overexpression recruits Atg14 and Uvrag to mitochondria. Miga-mediated stabilization of Syx17 (STX17) is dependent on Atg14, while PI3K activity enhancement by Miga requires Uvrag. |
Co-immunoprecipitation of Miga with Atg14/Uvrag, gain-of-function overexpression localization assay, genetic loss-of-function in Drosophila, PI3P measurement, Syx17 stability assay |
Cell reports |
Medium |
36323251
|
| 2023 |
MARCH7 (MARCHF7), an E3 ubiquitin ligase, ubiquitinates ATG14 with K6-, K11-, and K63-linked mixed polyubiquitin chains, causing ATG14 aggregation and reduced solubility. Ubiquitinated ATG14 has fewer interactions with STX17, inhibiting autophagy flux. MARCH7 depletion decreases aggresome-like induced structures. |
E3 ligase identification by Co-IP, in vitro/in cell ubiquitination assay with chain-type mapping, ATG14 solubility fractionation, STX17 binding assay for ubiquitinated vs. unmodified ATG14, MARCH7 KO cell analysis |
Cell reports |
High |
37632749
|
| 2023 |
RUNDC1 interacts with ATG14 and negatively regulates autophagy by blocking autophagosome-lysosome fusion. RUNDC1 clasps the ATG14-STX17-SNAP29 complex by stimulating ATG14 homo-oligomerization, preventing ATG14 dissociation and VAMP8 binding to STX17-SNAP29. Phosphorylation of RUNDC1 Ser379 is crucial for this inhibitory mechanism in human cells and zebrafish. |
Co-immunoprecipitation, gain/loss-of-function in human cells and zebrafish, STX17-SNAP29-VAMP8 complex assembly assay, ATG14 homo-oligomerization assay, phosphorylation site mutagenesis |
Cell death and differentiation |
Medium |
37684417
|
| 2023 |
USP1 interacts with ATG14, deubiquitinates it, and enhances ATG14 protein stability by reducing ubiquitin binding. USP1 inhibition promotes proteasome-dependent ATG14 degradation, reducing autophagy initiation and flux. |
Co-immunoprecipitation of USP1 with ATG14, ubiquitination level assay with USP1 overexpression/inhibition, proteasomal inhibition rescue, autophagy flux measurement |
The Journal of biological chemistry |
Medium |
39814232
|
| 2023 |
S. pneumoniae CbpC interacts with ATG14 via ATG14's coiled-coil domain and CbpC residue Y83 (dp3 domain), activating autophagy. However, CbpC also acts as a decoy by recruiting ATG14 to p62/SQSTM1-dependent autophagic degradation, depleting ATG14 and impairing ATG14-STX17 complex formation, thereby suppressing bactericidal autophagy and promoting pneumococcal intracellular survival. |
Co-immunoprecipitation of CbpC with ATG14, domain mutagenesis (Y83), ATG14 protein level measurement in infected cells, ATG14-STX17 complex formation assay, bacterial survival assay |
EMBO reports |
Medium |
32239622
|
| 2023 |
ATG14 and ATG7 are both required to restrict Mycobacterium tuberculosis replication in human iPSC-derived macrophages. ATG14 deletion specifically impairs phagosome-lysosome fusion in phagosomes containing Mtb, enabling both cytosolic and phagosomal bacterial restriction, while ATG7 primarily controls canonical autophagy of cytosolic Mtb. |
CRISPR-Cas9 deletion of ATG14 and ATG7 in iPSDM, single-cell high-content imaging with Mtb replication reporters, phagosome-lysosome fusion quantitative imaging with Mtb mutant panel |
Nature microbiology |
High |
36959508
|
| 2024 |
ATG14 targets lipid droplets (LDs) via its BATS domain and acts as an autophagic receptor for 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 and induce lipophagy to degrade the anti-viral protein Viperin. |
BATS domain mutagenesis for LD targeting, Co-immunoprecipitation of ATG14 with ATG8 family members and STX18, STX18 knockdown-dependent lipophagy assay, Viperin degradation assay with coronavirus M protein |
Nature communications |
High |
38245527
|
| 2024 |
ATG14 localizes to lipid droplets via its BATS domain and directly interacts with adipose triglyceride lipase (ATGL) and its coactivator CGI-58. ATG14 enhances the ATGL-CGI58 interaction; ATG14 deficiency markedly decreases triglyceride hydrolysis in hepatocytes. |
Fluorescence imaging of ATG14 on lipid droplets, BATS domain deletion/mutagenesis, co-immunoprecipitation of ATG14 with ATGL and CGI-58, in vitro lipolysis assay in ATG14-deficient hepatocytes and mouse livers |
Metabolism: clinical and experimental |
High |
37741434
|
| 2024 |
GULP1 (engulfment adaptor GULP1) interacts with ATG14 and potentiates its stimulatory effect on PI3KC3-C1 activity. GULP1 facilitates targeting of ATG14 to the ER. A GULP1 mutation disrupting the GULP1-ATG14 interaction abolishes these effects. GULP1-APP complex (via GULP1-ATG14 axis) further enhances PI3KC3-C1 activity and promotes APP processing by directing APP into autophagic vacuoles. |
Co-immunoprecipitation of GULP1 with ATG14, PI3KC3-C1 kinase activity assay, GULP1 mutant (disrupted ATG14 binding) rescue assay, ER targeting assay by fluorescence microscopy, autophagic vacuole fractionation for APP/ATG14 co-localization |
Cellular and molecular life sciences |
Medium |
39080084
|
| 2012 |
ATG14 is regulated transcriptionally by FoxO transcription factors and circadian clock machinery (Clock/Bmal1) in mouse liver. Knockdown of ATG14 in mouse liver leads to elevated triglycerides in liver and serum; overexpression of ATG14 improves hypertriglyceridemia in high-fat diet and FoxO1/3/4 liver KO mice. |
Luciferase reporter assays, chromatin immunoprecipitation (ChIP) for FoxOs and Clock/Bmal1 at ATG14 promoter, liver-specific ATG14 knockdown and overexpression in mice, liver triglyceride and serum lipid measurement |
The Journal of biological chemistry |
Medium |
22992773
|
| 2025 |
ATG14 safeguards oviduct cellular integrity by suppressing pyroptosis. Conditional depletion of Atg14 in oviduct causes severe structural abnormalities and abnormal embryo retention. Mechanistically, Atg14 loss triggers unscheduled pyroptosis via altered mitochondrial integrity in non-ciliary oviduct cells; pharmacological pyroptosis activation phenocopies the genetic defect. |
Conditional oviduct-specific Atg14 KO mice, histopathology for structural defects, embryo tracking assay, pyroptosis marker analysis, mitochondrial integrity assay, pharmacological pyroptosis induction |
eLife |
Medium |
40100261
|
| 2013 |
CCCP-induced LC3 lipidation is independent of Beclin 1 and ATG14; FIP200 and ATG13 (ULK complex components) are only partially required, while ATG9 is required for CCCP-induced LC3 lipidation. This establishes that CCCP/mitophagy-induced autophagy can bypass the Beclin 1/ATG14 nucleation complex. |
Genetic deletion/siRNA knockdown of Beclin 1, ATG14, FIP200, ATG13, ATG9; LC3 lipidation assay (Western blot) under CCCP treatment |
Biochemical and biophysical research communications |
Medium |
23402761
|
| 2014 |
Dapper1 (Dpr1) directly interacts with both Beclin1 and Atg14L, enhances the Beclin1-Vps34 interaction, and increases Vps34 lipid kinase activity, promoting autophagosome formation. Dpr1 ablation in the CNS results in motor coordination defects and accumulation of p62 and ubiquitinated proteins. |
Co-immunoprecipitation of Dpr1 with Beclin1 and Atg14L, Vps34 kinase activity assay, Dpr1 CNS-specific KO mice, LC3 puncta formation assay, p62/ubiquitin accumulation |
Cell research |
Medium |
24980960
|