RUBCN encodes Rubicon (RUN domain Beclin-1-interacting and cysteine-rich domain-containing protein), a negative regulator of late-stage autophagy and endolysosomal maturation. Its middle PI3K-binding region associates with the UVRAG-containing class III phosphatidylinositol 3-kinase complex II (Beclin 1-VPS34-VPS15-UVRAG), contacting the Beclin 1 BARA domain, and inhibits VPS34 lipid kinase activity; its C-terminal zinc-cluster Rubicon homology (RH) domain binds GTP-bound Rab7, targeting Rubicon to late endosomes and lysosomes. By restraining PI3KC3-C2 and sequestering UVRAG from the HOPS tethering complex, Rubicon slows autophagosome maturation, autophagosome-lysosome fusion and endosome maturation. Rubicon expression rises with age and under lipid overload, and its loss enhances autophagic flux. In phagocytes Rubicon is also linked to NADPH oxidase-dependent phagosome functions and to feedback inhibition of CARD9 signalling downstream of pattern-recognition receptors.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005769 early endosome | IBA GO_REF:0000033 | ACCEPT | Summary: early endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0005770 late endosome | IBA GO_REF:0000033 | ACCEPT | Summary: late endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0045806 negative regulation of endocytosis | IBA GO_REF:0000033 | ACCEPT | Summary: Negative regulation of endocytosis/endocytic trafficking is a core Rubicon function. Reason: Rubicon inhibits endosome maturation and endocytic pathway progression through Rab7 and UVRAG/PI3KC3-C2 interactions. Supporting Evidence: PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS PMID:20943950 specifically and directly interact with Rab7 via their RH domain PMID:20943950 simultaneously binding both Rab7 and PI3-kinase PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes |
| GO:1901097 negative regulation of autophagosome maturation | IBA GO_REF:0000033 | ACCEPT | Summary: Negative regulation of autophagosome maturation is the central RUBCN autophagy annotation. Reason: Rubicon inhibits UVRAG-containing PI3KC3-C2 and autophagosome maturation; knockdown enhances autophagy at the maturation step. Supporting Evidence: PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes PMID:19270696 Knockdown of Rubicon caused enhancement of autophagy, especially at the maturation step PMID:21062745 negative regulator of PI3KC3 and autophagosome maturation file:human/RUBCN/RUBCN-uniprot.txt negatively regulates PI3K complex II (PI3KC3-C2) function in autophagy PMID:21062745 interacts with the PI3KC3 catalytic subunit hVps34 via its RUN domain PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon file:human/RUBCN/RUBCN-uniprot.txt Inhibits PIK3C3 activity |
| GO:1901981 phosphatidylinositol phosphate binding | IBA GO_REF:0000033 | MARK AS OVER ANNOTATED | Summary: Phosphatidylinositol phosphate binding is inherited from a Eukaryota-root PAINT node seeded only by RUBCNL/Pacer; direct lipid binding has not been shown for Rubicon, and the available evidence argues against it. Reason: The only experimental seed for PTN002915557 is Pacer (Q9H714, IDA PMID:28306502). For Rubicon, a PtdIns(3)P bead pull-down was negative (PMID:19270693), and the RH-domain crystal structure shows that its FYVE-like zinc cluster lacks the basic residues needed for PI(3)P binding, the authors noting that Rubicon has not been reported to bind phospholipids (PMID:32632011). Rubicon membrane targeting is explained by binding to Rab7-GTP and PI3KC3-C2 instead. Lipid binding appears to be a Pacer-lineage property, not an established ancestral function. Propagation Review Root cause: PROPAGATION BAD Failure modes: FUNCTIONAL DIVERGENCE Sources checked: PANTHER:PTN002915557 · PTN002915557 SUPPORTS SOURCE BUT NOT TARGET Eukaryota-root node seeded only by RUBCNL/Pacer lipid-strip binding (PMID:28306502); Rubicon was PtdIns(3)P-bead negative (PMID:19270693) and lacks FYVE basic residues (PMID:32632011). Supporting Evidence: PMID:19270693 Rubicon-EGFP was not pulled down by PtdIns(3)P-conjugated sepharose beads PMID:32632011 Rubicon has not been reported to bind specifically to phospholipids PMID:32632011 However, the Rubicon Zn cluster 1 lacks some of the basic residues required for canonical FYVE domain phosphatidylinositol 3-phosphate (PI[3]P) binding |
| GO:0005764 lysosome | IEA GO_REF:0000044 | ACCEPT | Summary: Lysosome localization is supported and relevant to late endosome/autophagosome maturation. Reason: Rubicon is predominantly in late endosome/lysosome compartments and regulates delivery/fusion steps toward lysosomes. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Lysosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG |
| GO:0005769 early endosome | IEA GO_REF:0000044 | ACCEPT | Summary: early endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0005770 late endosome | IEA GO_REF:0000044 | ACCEPT | Summary: late endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0005515 protein binding | IPI PMID:20562859 Network organization of the human autophagy system. | REMOVE | Summary: Generic protein binding to BECN1 from an autophagy-network proteomics screen conveys no function of RUBCN. Reason: The BECN1 interaction is consistent with Rubicon association with PI3KC3-C2, but an autophagy-network proteomics screen reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:20562859 KIAA0831/ATG14 and KIAA0226/RUBICON, were found to interact with BECN1 and PIK3C3 |
| GO:0005515 protein binding | IPI PMID:21062745 The RUN domain of rubicon is important for hVps34 binding, l... | MODIFY | Summary: Rubicon binding to BECN1 reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:21062745 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the BECN1 interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005515 protein binding | IPI PMID:22493499 Receptor signaling lymphocyte-activation molecule family 1 (... | REMOVE | Summary: Generic protein binding to BECN1 from a Slamf1/NOX2 phagosome study conveys no function of RUBCN. Reason: The BECN1 interaction is consistent with Rubicon association with PI3KC3-C2, but a Slamf1/NOX2 phagosome study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:22493499 we transfected HEK293 cells with Slamf1, Vps34/15, and Beclin-1 together with HA-UVRAG, Atg14L-EGFP, or EGFP-Rubicon |
| GO:0005515 protein binding | IPI PMID:23954414 Beclin 2 functions in autophagy, degradation of G protein-co... | REMOVE | Summary: Generic protein binding to BECN1 from a Beclin 2 study conveys no function of RUBCN. Reason: The BECN1 interaction is consistent with Rubicon association with PI3KC3-C2, but a Beclin 2 study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:23954414 No interaction was detected between mouse or human Beclin 2 and the Beclin 1-binding negative regulator of autophagy, Rubicon. |
| GO:0005515 protein binding | IPI PMID:24034250 EGFR-mediated Beclin 1 phosphorylation in autophagy suppress... | REMOVE | Summary: Generic protein binding to EGFR from an EGFR-Beclin 1 phosphorylation study conveys no function of RUBCN. Reason: The EGFR interaction is consistent with Rubicon association with PI3KC3-C2 (or, for EGFR, with EGFR-mediated release of Beclin 1 from Rubicon), but an EGFR-Beclin 1 phosphorylation study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:24034250 did not disrupt Beclin 1 EEE/EGFR, Beclin 1 EEE/Rubicon, or Beclin 1/Bcl-2 interactions |
| GO:0005515 protein binding | IPI PMID:25594178 A kinase-independent role for EGF receptor in autophagy init... | REMOVE | Summary: Generic protein binding to EGFR from an EGFR/LAPTM4B/Sec5 autophagy-initiation study conveys no function of RUBCN. Reason: The EGFR interaction is consistent with Rubicon association with PI3KC3-C2 (or, for EGFR, with EGFR-mediated release of Beclin 1 from Rubicon), but an EGFR/LAPTM4B/Sec5 autophagy-initiation study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:25594178 LAPTM4B and Sec5 promote EGFR association with the autophagy inhibitor Rubicon, which in turn disassociates Beclin 1 from Rubicon to initiate autophagy. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to BRD1 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: BRD1 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: Generic protein binding to BECN1 from the BioPlex affinity-purification mass spectrometry interactome conveys no function of RUBCN. Reason: The BECN1 interaction is consistent with Rubicon association with PI3KC3-C2, but the BioPlex affinity-purification mass spectrometry interactome reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:33961781 Here we present BioPlex 3.0, the most complete model of the human interactome to date |
| GO:0005515 protein binding | IPI PMID:34386498 ORF3a-Mediated Incomplete Autophagy Facilitates Severe Acute... | REMOVE | Summary: Generic protein binding to UVRAG from a SARS-CoV-2 ORF3a study conveys no function of RUBCN. Reason: The UVRAG interaction is consistent with Rubicon association with PI3KC3-C2, but a SARS-CoV-2 ORF3a study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:34386498 the presence of ORF3a significantly reduced the interaction between UVRAG and Beclin 1 or other Beclin 1 complex components, such as Vps34 and Rubicon |
| GO:0005515 protein binding | IPI PMID:35271311 OpenCell: Endogenous tagging for the cartography of human ce... | REMOVE | Summary: Generic protein binding to PIK3C3 from the OpenCell endogenous-tagging interactome conveys no function of RUBCN. Reason: The PIK3C3 interaction is consistent with Rubicon association with PI3KC3-C2, but the OpenCell endogenous-tagging interactome reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:35271311 We combined genome engineering, confocal live-cell imaging, mass spectrometry and data science to systematically map the localization and interactions of human proteins. |
| GO:0005515 protein binding | IPI PMID:35512704 Systematic discovery of mutation-directed neo-protein-protei... | REMOVE | Summary: Generic protein binding to EGFR from a cancer neo-interaction screen conveys no function of RUBCN. Reason: The EGFR interaction is consistent with Rubicon association with PI3KC3-C2 (or, for EGFR, with EGFR-mediated release of Beclin 1 from Rubicon), but a cancer neo-interaction screen reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:35512704 a BRET n technology-based differential PPI discovery platform that allows comparative screening of wild-type (WT) and MUT allele counterparts |
| GO:0005764 lysosome | EXP PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | ACCEPT | Summary: Lysosome localization is supported and relevant to late endosome/autophagosome maturation. Reason: Rubicon is predominantly in late endosome/lysosome compartments and regulates delivery/fusion steps toward lysosomes. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Lysosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG |
| GO:0005769 early endosome | EXP PMID:20974968 Rubicon controls endosome maturation as a Rab7 effector. | ACCEPT | Summary: early endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0051898 negative regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction | IDA PMID:21062745 The RUN domain of rubicon is important for hVps34 binding, l... | MODIFY | Summary: The PI3K/AKT pathway wording is misleading for Rubicon hVps34 inhibition. Reason: The cited evidence supports inhibition of class III PI3K/VPS34 lipid kinase activity and autophagosome maturation, not a general PI3K/AKT signaling pathway role. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity negative regulation of autophagosome maturation Supporting Evidence: PMID:21062745 interacts with the PI3KC3 catalytic subunit hVps34 via its RUN domain PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon file:human/RUBCN/RUBCN-uniprot.txt Inhibits PIK3C3 activity PMID:21062745 negative regulator of PI3KC3 and autophagosome maturation file:human/RUBCN/RUBCN-uniprot.txt negatively regulates PI3K complex II (PI3KC3-C2) function in autophagy |
| GO:0141039 phosphatidylinositol 3-kinase inhibitor activity | IDA PMID:21062745 The RUN domain of rubicon is important for hVps34 binding, l... | ACCEPT | Summary: PI3K inhibitor activity is the best molecular-function annotation for Rubicon. Reason: Rubicon inhibits PIK3C3/VPS34 lipid kinase activity through RUN-domain-dependent interaction with hVps34. Supporting Evidence: PMID:21062745 interacts with the PI3KC3 catalytic subunit hVps34 via its RUN domain PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon file:human/RUBCN/RUBCN-uniprot.txt Inhibits PIK3C3 activity |
| GO:1901097 negative regulation of autophagosome maturation | TAS PMID:21118109 The role of ESCRT proteins in fusion events involving lysoso... | ACCEPT | Summary: Negative regulation of autophagosome maturation is the central RUBCN autophagy annotation. Reason: Rubicon inhibits UVRAG-containing PI3KC3-C2 and autophagosome maturation; knockdown enhances autophagy at the maturation step. Supporting Evidence: PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes PMID:19270696 Knockdown of Rubicon caused enhancement of autophagy, especially at the maturation step PMID:21062745 negative regulator of PI3KC3 and autophagosome maturation file:human/RUBCN/RUBCN-uniprot.txt negatively regulates PI3K complex II (PI3KC3-C2) function in autophagy PMID:21062745 interacts with the PI3KC3 catalytic subunit hVps34 via its RUN domain PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon file:human/RUBCN/RUBCN-uniprot.txt Inhibits PIK3C3 activity |
| GO:0071985 multivesicular body sorting pathway | TAS PMID:21118109 The role of ESCRT proteins in fusion events involving lysoso... | KEEP AS NON CORE | Summary: Multivesicular body sorting is supported as endosomal trafficking context but is not the core autophagy-regulation term. Reason: Rubicon negatively regulates endocytic pathway progression and endosome maturation; retain this as a non-core endosomal trafficking branch. Supporting Evidence: PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS PMID:20943950 specifically and directly interact with Rab7 via their RH domain PMID:20943950 simultaneously binding both Rab7 and PI3-kinase |
| GO:1901097 negative regulation of autophagosome maturation | IMP PMID:21062745 The RUN domain of rubicon is important for hVps34 binding, l... | ACCEPT | Summary: Negative regulation of autophagosome maturation is the central RUBCN autophagy annotation. Reason: Rubicon inhibits UVRAG-containing PI3KC3-C2 and autophagosome maturation; knockdown enhances autophagy at the maturation step. Supporting Evidence: PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes PMID:19270696 Knockdown of Rubicon caused enhancement of autophagy, especially at the maturation step PMID:21062745 negative regulator of PI3KC3 and autophagosome maturation file:human/RUBCN/RUBCN-uniprot.txt negatively regulates PI3K complex II (PI3KC3-C2) function in autophagy PMID:21062745 interacts with the PI3KC3 catalytic subunit hVps34 via its RUN domain PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon file:human/RUBCN/RUBCN-uniprot.txt Inhibits PIK3C3 activity |
| GO:0005515 protein binding | IPI PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | MODIFY | Summary: Rubicon binding to BECN1 reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:19270696 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the BECN1 interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005769 early endosome | IDA PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | ACCEPT | Summary: early endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0005770 late endosome | IDA PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | ACCEPT | Summary: late endosome localization is supported for Rubicon endosome maturation biology. Reason: RUBCN is an endosomal/lysosomal Rab7 and UVRAG/PI3KC3-C2 regulator; early/late endosome localization fits the core negative endosome/autophagosome maturation role. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt Predominantly located in late endosomes/lysosomes file:human/RUBCN/RUBCN-uniprot.txt Late endosome {ECO:0000269|PubMed:19270696} file:human/RUBCN/RUBCN-uniprot.txt Early endosome PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS |
| GO:0005794 Golgi apparatus | IDA NOT PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | ACCEPT | Summary: The negated Golgi colocalization annotation is supported by curated localization evidence. Reason: This is a NOT annotation. Primary Rubicon localization is endosome/lysosome, and UniProt explicitly notes that Rubicon is not detected in the Golgi apparatus. Supporting Evidence: file:human/RUBCN/RUBCN-uniprot.txt not at all in the Golgi apparatus PMID:19270696 GFP-Rubicon was localized at the endosome/lysosome |
| GO:0010507 negative regulation of autophagy | IMP PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | ACCEPT | Summary: Negative regulation of autophagy is supported and core, though less specific than autophagosome maturation. Reason: Rubicon knockdown enhances autophagy and maturation; the specific mechanism is PI3KC3-C2 inhibition and impaired autophagosome maturation. Supporting Evidence: PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes PMID:19270696 Knockdown of Rubicon caused enhancement of autophagy, especially at the maturation step PMID:21062745 negative regulator of PI3KC3 and autophagosome maturation file:human/RUBCN/RUBCN-uniprot.txt negatively regulates PI3K complex II (PI3KC3-C2) function in autophagy PMID:21062745 interacts with the PI3KC3 catalytic subunit hVps34 via its RUN domain PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon file:human/RUBCN/RUBCN-uniprot.txt Inhibits PIK3C3 activity |
| GO:0045806 negative regulation of endocytosis | IMP PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | ACCEPT | Summary: Negative regulation of endocytosis/endocytic trafficking is a core Rubicon function. Reason: Rubicon inhibits endosome maturation and endocytic pathway progression through Rab7 and UVRAG/PI3KC3-C2 interactions. Supporting Evidence: PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG PMID:20974968 sequesters UVRAG from C-VPS/HOPS PMID:20943950 specifically and directly interact with Rab7 via their RH domain PMID:20943950 simultaneously binding both Rab7 and PI3-kinase PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes |
| GO:0034272 phosphatidylinositol 3-kinase complex, class III, type II | IPI PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | NEW | Summary: Rubicon should be added as a PI3KC3-C2-associated regulatory subunit/context component. Reason: RUBCN associates with the UVRAG-containing class III PI3K complex II and directly interacts with PIK3C3 and UVRAG; this is the specific complex context missing from GOA. Supporting Evidence: PMID:19270696 Rubicon binds only to a subpopulation of UVRAG complexes file:human/RUBCN/RUBCN-uniprot.txt Associates with PI3K (PI3KC3/PI3K-III/class III file:human/RUBCN/RUBCN-uniprot.txt complex II (PI3KC3-C2) |
| GO:0005515 protein binding | IPI PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | MODIFY | Summary: Rubicon binding to PIK3C3 reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:19270696 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the PIK3C3 interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005515 protein binding | IPI PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | MODIFY | Summary: Rubicon binding to PIK3R4 reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:19270696 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the PIK3R4 interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005515 protein binding | IPI PMID:19270696 Two Beclin 1-binding proteins, Atg14L and Rubicon, reciproca... | MODIFY | Summary: Rubicon binding to UVRAG reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:19270696 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the UVRAG interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005515 protein binding | IPI PMID:20562859 Network organization of the human autophagy system. | REMOVE | Summary: Generic protein binding to PIK3C3 from an autophagy-network proteomics screen conveys no function of RUBCN. Reason: The PIK3C3 interaction is consistent with Rubicon association with PI3KC3-C2, but an autophagy-network proteomics screen reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:20562859 KIAA0831/ATG14 and KIAA0226/RUBICON, were found to interact with BECN1 and PIK3C3 |
| GO:0005515 protein binding | IPI PMID:20562859 Network organization of the human autophagy system. | REMOVE | Summary: Generic protein binding to UVRAG from an autophagy-network proteomics screen conveys no function of RUBCN. Reason: The UVRAG interaction is consistent with Rubicon association with PI3KC3-C2, but an autophagy-network proteomics screen reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:20562859 KIAA0831/ATG14 and KIAA0226/RUBICON, were found to interact with BECN1 and PIK3C3 |
| GO:0005515 protein binding | IPI PMID:21062745 The RUN domain of rubicon is important for hVps34 binding, l... | MODIFY | Summary: Rubicon binding to PIK3C3 reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:21062745 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the PIK3C3 interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005515 protein binding | IPI PMID:21062745 The RUN domain of rubicon is important for hVps34 binding, l... | MODIFY | Summary: Rubicon binding to UVRAG reflects its association with the UVRAG-containing class III PI3K complex II, which it inhibits; generic protein binding should be replaced by the informative activity. Reason: PMID:21062745 characterizes Rubicon as a Beclin 1/UVRAG/VPS34 complex-associated negative regulator; the functional content of the UVRAG interaction is inhibition of PI3KC3 lipid kinase activity (PMID:21062745, PMID:19270693). The binding itself is a means to that activity, so the replacement is phosphatidylinositol 3-kinase inhibitor activity. Proposed replacements: phosphatidylinositol 3-kinase inhibitor activity Supporting Evidence: PMID:21062745 inhibition of PI3KC3 lipid kinase activity by Rubicon PMID:19270693 Rubicon reduces Vps34 activity and downregulates autophagy file:human/RUBCN/RUBCN-uniprot.txt interacts directly with PI3KC3 and UVRAG |
| GO:0005515 protein binding | IPI PMID:22493499 Receptor signaling lymphocyte-activation molecule family 1 (... | REMOVE | Summary: Generic protein binding to UVRAG from a Slamf1/NOX2 phagosome study conveys no function of RUBCN. Reason: The UVRAG interaction is consistent with Rubicon association with PI3KC3-C2, but a Slamf1/NOX2 phagosome study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:22493499 we transfected HEK293 cells with Slamf1, Vps34/15, and Beclin-1 together with HA-UVRAG, Atg14L-EGFP, or EGFP-Rubicon |
| GO:0005515 protein binding | IPI PMID:24034250 EGFR-mediated Beclin 1 phosphorylation in autophagy suppress... | REMOVE | Summary: Generic protein binding to BECN1 from an EGFR-Beclin 1 phosphorylation study conveys no function of RUBCN. Reason: The BECN1 interaction is consistent with Rubicon association with PI3KC3-C2, but an EGFR-Beclin 1 phosphorylation study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:24034250 did not disrupt Beclin 1 EEE/EGFR, Beclin 1 EEE/Rubicon, or Beclin 1/Bcl-2 interactions |
| GO:0005515 protein binding | IPI PMID:25594178 A kinase-independent role for EGF receptor in autophagy init... | REMOVE | Summary: Generic protein binding to BECN1 from an EGFR/LAPTM4B/Sec5 autophagy-initiation study conveys no function of RUBCN. Reason: The BECN1 interaction is consistent with Rubicon association with PI3KC3-C2, but an EGFR/LAPTM4B/Sec5 autophagy-initiation study reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:25594178 LAPTM4B and Sec5 promote EGFR association with the autophagy inhibitor Rubicon, which in turn disassociates Beclin 1 from Rubicon to initiate autophagy. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to DRC4 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: DRC4 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to ZNF250 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: ZNF250 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to BYSL from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: BYSL was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to VEZF1 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: VEZF1 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to FAM161A from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: FAM161A was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to ZNF648 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: ZNF648 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to FAM9B from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: FAM9B was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to TCEANC from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: TCEANC was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to DPF2 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: DPF2 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to ENKD1 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: ENKD1 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to NME7 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: NME7 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Generic protein binding to ANKRD11 from the HuRI binary yeast two-hybrid interactome screen conveys no function of RUBCN. Reason: ANKRD11 was detected as a Rubicon partner only in the HuRI binary yeast two-hybrid interactome screen; there is no functional follow-up linking this interaction to Rubicon biology, so no informative molecular function can be substituted. Removal does not imply the interaction is false. Supporting Evidence: PMID:32296183 yeast two-hybrid (Y2H) represents the only binary PPI assay that can be operated at sufficient throughput to systematically screen the human proteome for binary PPIs |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: Generic protein binding to PIK3C3 from the BioPlex affinity-purification mass spectrometry interactome conveys no function of RUBCN. Reason: The PIK3C3 interaction is consistent with Rubicon association with PI3KC3-C2, but the BioPlex affinity-purification mass spectrometry interactome reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:33961781 Here we present BioPlex 3.0, the most complete model of the human interactome to date |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: Generic protein binding to UVRAG from the BioPlex affinity-purification mass spectrometry interactome conveys no function of RUBCN. Reason: The UVRAG interaction is consistent with Rubicon association with PI3KC3-C2, but the BioPlex affinity-purification mass spectrometry interactome reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:33961781 Here we present BioPlex 3.0, the most complete model of the human interactome to date |
| GO:0005515 protein binding | IPI PMID:35271311 OpenCell: Endogenous tagging for the cartography of human ce... | REMOVE | Summary: Generic protein binding to UVRAG from the OpenCell endogenous-tagging interactome conveys no function of RUBCN. Reason: The UVRAG interaction is consistent with Rubicon association with PI3KC3-C2, but the OpenCell endogenous-tagging interactome reports the interaction without a functional assay of Rubicon activity. The informative activity (GO:0141039 phosphatidylinositol 3-kinase inhibitor activity) is already captured from PMID:21062745. Removal does not imply the interaction is false. Supporting Evidence: PMID:35271311 We combined genome engineering, confocal live-cell imaging, mass spectrometry and data science to systematically map the localization and interactions of human proteins. |
| GO:0031267 small GTPase binding | IDA PMID:32632011 Structural basis for autophagy inhibition by the human Rubic... | NEW | Summary: The Rubicon RH domain binds GTP-bound Rab7 directly; this targets Rubicon to late endosomes and is required for its autophagy-inhibitory function. Reason: Direct binding of the RH domain to Rab7-GTP is shown biochemically (PMID:20943950), by a 2.8 A co-crystal structure, and by interface mutants that abolish colocalization and relieve autophagy inhibition (PMID:32632011). Rab7 binding is an activity of Rubicon itself, not just a requirement. No GOA row currently captures it (only GO:0005515). Supporting Evidence: PMID:32632011 Rubicon-mediated inhibition of autophagy requires the interaction of the C-terminal Rubicon homology (RH) domain of Rubicon with Rab7-GTP PMID:20943950 specifically and directly interact with Rab7 via their RH domain PMID:20974968 prevents endosome maturation through differential interactions with Rab7 and UVRAG |
Loading supporting content…
Download this section (compressed HTML)Q: Should RUBCN be curated as part_of GO:0034272 PI3KC3-C2, or represented only by PI3K inhibitor activity and UVRAG/Rab7 interaction context?
Suggested experts: GO autophagy editors, ComplexPortal curators
Q: Should negative regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction be replaced in GOA by phosphatidylinositol 3-kinase inhibitor activity for RUBCN?
Suggested experts: GO molecular function editors, GO signaling editors
Q: Is GO:1901981 phosphatidylinositol phosphate binding at the Eukaryota root node PTN002915557 an ancestral family property or a Pacer/RUBCNL-specific gain? Rubicon was negative in a PtdIns(3)P bead pull-down and its FYVE-like zinc cluster lacks canonical basic residues.
Suggested experts: PAINT curators
Experiment: Use RUBCN knockout/rescue with RUN-domain hVps34-binding mutants and RH-domain Rab7-binding mutants to measure PI3KC3 lipid kinase activity, Rab7 activation, EGFR/endosome maturation, and autophagosome-lysosome fusion.
Hypothesis: RUBCN requires both PI3KC3-C2 and Rab7/UVRAG interaction surfaces to inhibit autophagosome and endosome maturation.
Type: Rubicon PI3KC3-C2/Rab7 separation-of-function rescue
Experiment: Perform live-cell imaging of tagged endogenous RUBCN, UVRAG, Rab7, and HOPS markers during starvation and endocytic cargo maturation to test whether RUBCN leaves endosomal/autophagosomal sites before fusion competence.
Hypothesis: RUBCN occupancy marks inhibited PI3KC3-C2/Rab7 compartments and must be relieved for maturation/fusion progression.
Type: endogenous live-cell maturation imaging
Loading supporting content…
Download this section (compressed HTML)Loading supporting content…
Download this section (compressed HTML)Loading supporting content…
Download this section (compressed HTML)