Affinage

RAB11FIP3

Rab11 family-interacting protein 3 · UniProt O75154

Length
756 aa
Mass
82.4 kDa
Annotated
2026-06-10
15 papers in source corpus 12 papers cited in narrative 12 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RAB11FIP3 (Eferin/Rab11-FIP3) is a Rab11/Arf6 dual-effector scaffolding protein that couples Rab11-positive recycling endosomes to the cytoplasmic dynein motor to organize the pericentrosomal endosomal recycling compartment (ERC) and to direct endosome-driven processes including cytokinesis and immune-synapse signaling (PMID:11481332, PMID:17394487, PMID:20026645). A C-terminal 20-residue domain mediates direct Rab11 binding, and this interaction is required for correct effector localization to a Rab11-positive pericentrosomal compartment (PMID:11481332, PMID:15158446). RAB11FIP3 is essential for ERC structural integrity: loss of RAB11FIP3 or Rab11-binding-deficient mutants disperses ERC markers and blocks transferrin delivery to the pericentrosomal region, a function that depends on a dimerizing coiled-coil region (residues 463–692) (PMID:17394487). It establishes minus-end-directed transport by directly binding cytoplasmic dynein light intermediate chains DLIC-1 (DYNC1LI1) and DLIC-2 (DYNC1LI2), forming Rab11a–FIP3–DLIC ternary complexes and recruiting dynein onto endosomal membranes (PMID:20026645, PMID:20214888). In parallel, RAB11FIP3 preferentially binds the Arf6 C-terminus and acts as a scaffold delivering Arf6 to the cleavage furrow and midbody during cytokinesis, with sequential Rab11- then Arf6-dependent targeting of FIP3 vesicles to the Flemming body during abscission (PMID:17628206, PMID:21790911). Its activity is cell-cycle gated by Cdk1–cyclin B phosphorylation at S-102, which is high in metaphase and removed at telophase as FIP3 translocates from cytosol to membranes (PMID:22401586). Through Rab11-dependent endosomal recycling, RAB11FIP3 targets Rac1 and Lck to the immunological synapse to control actin dynamics, TCR proximal signaling and T-cell activation (PMID:27154205, PMID:28235866), and drives endosomal recycling of PD-L1 to the cancer-cell surface to promote tumor immune evasion (PMID:38211652).

Mechanistic history

Synthesis pass · year-by-year structured walk · 12 steps
  1. 2001 High

    Established RAB11FIP3 as a Rab11 effector and defined the minimal binding determinant, answering how the protein is recruited to recycling-endosome machinery.

    Evidence In vitro Rab11 binding assays with C-terminal domain mutagenesis and in vivo localization across the Rab11/25-interacting protein family

    PMID:11481332

    Open questions at the time
    • Did not define downstream effector function
    • No structural model of the Rab11-binding interface
  2. 2004 Medium

    Placed RAB11FIP3 at a pericentrosomal Rab11 compartment in interphase and the cleavage furrow in cytokinesis, linking it to cytoskeleton-dependent positioning rather than bulk transferrin recycling.

    Evidence Immunofluorescence with cytoskeletal disruption and N-terminal truncation mutant; transferrin recycling assay

    PMID:15158446

    Open questions at the time
    • Mechanism linking FIP3 to microtubule/actin not identified
    • Truncation result based on single dominant-negative construct
  3. 2007 High

    Demonstrated RAB11FIP3 is required for ERC structural integrity and mapped a dimerizing coiled-coil domain as the functional element, defining its scaffolding role at the recycling compartment.

    Evidence siRNA knockdown, Rab11-binding-deficient mutant, transferrin trafficking, and biochemical coiled-coil dimerization analysis

    PMID:17394487

    Open questions at the time
    • Did not identify the motor or partners driving pericentrosomal accumulation
    • Single-lab phenotype
  4. 2007 High

    Identified Arf6 as a second effector partner and showed FIP3 scaffolds Arf6 recruitment to the furrow and midbody, connecting FIP3 to cytokinetic membrane delivery.

    Evidence Biochemical affinity comparison among Arf GTPases, Co-IP, domain mapping, and live-cell cytokinesis imaging

    PMID:17628206

    Open questions at the time
    • Functional consequence of Arf6 binding for abscission completion not quantified here
    • Did not resolve coordination with Rab11 binding
  5. 2009 High

    Resolved the motor link by showing direct FIP3–DLIC-1 binding and Rab11a–FIP3–DLIC-1 ternary complex formation, explaining minus-end-directed transport of recycling endosomes to the ERC.

    Evidence Reciprocal Co-IP, GST pulldown, membrane recruitment, DLIC-1 knockdown, and truncation mutant disrupting ERC accumulation

    PMID:20026645

    Open questions at the time
    • Did not address whether DLIC-2 contributes
    • Regulation of motor engagement timing unresolved
  6. 2009 Medium

    Extended FIP3 function to cell migration, showing it controls Rac1-dependent actin dynamics and Arf6 localization at the leading edge of carcinoma cells.

    Evidence siRNA knockdown with motility, actin dynamics, and Arf6 localization readouts in MDA-MB-231 cells

    PMID:19327867

    Open questions at the time
    • Direct vs indirect control of Rac1 not distinguished here
    • Single cell-line model
  7. 2010 Medium

    Showed FIP3 also binds DLIC-2 via its N-terminal region, broadening the dynein-coupling repertoire and demonstrating dynein-sequestering effects on Golgi morphology.

    Evidence Co-IP with domain mapping, membrane recruitment, DLIC-2 knockdown transferrin assay, and overexpression Golgi morphology assessment

    PMID:20214888

    Open questions at the time
    • Relative roles of DLIC-1 vs DLIC-2 not delineated
    • Golgi fragmentation may be an overexpression artifact
  8. 2011 Medium

    Established the temporal logic of cytokinetic targeting: Rab11 governs endosome recruitment and transport while Arf6 governs midbody targeting during abscission.

    Evidence Endogenous-protein immunofluorescence and time-lapse imaging of FIP3, Rab11, and Arf6 through cytokinesis

    PMID:21790911

    Open questions at the time
    • Molecular trigger of the Rab11-to-Arf6 handoff unknown
    • Fate of Flemming body remnant function not addressed
  9. 2012 Medium

    Identified FIP3 as a cell-cycle-regulated phosphoprotein with Cdk1-cyclin B phosphorylating S-102, linking mitotic kinase activity to FIP3 cytosol-to-membrane translocation.

    Evidence Mass spectrometry phosphosite mapping, in vitro Cdk1-cyclin B kinase assay, phospho-staging, and phosphomimetic mutant binucleation scoring

    PMID:22401586

    Open questions at the time
    • Identified sites do not control spatial endosome distribution
    • Phosphatase responsible for telophase dephosphorylation unknown
  10. 2016 Medium

    Demonstrated FIP3 mediates Rac1 association with Rab11 endosomes and its delivery to the immunological synapse, controlling T-cell spreading, synapse symmetry, and activation.

    Evidence FIP3 knockdown/overexpression, Rac1 co-localization, live imaging, and Rac1-dependent T-cell cytokine assays

    PMID:27154205

    Open questions at the time
    • Direct FIP3–Rac1 binding vs endosome co-trafficking not separated
    • Single-lab T-cell model
  11. 2017 Medium

    Showed FIP3-dependent endosomal trafficking of Lck controls proximal TCR signaling and IL-2 expression, extending FIP3's synapse role to kinase delivery.

    Evidence FIP3 KD/OE with Rab11-interaction mutants, Lck localization, phosphotyrosine blotting, calcium flux, and IL-2 reporter assays

    PMID:28235866

    Open questions at the time
    • Whether Lck is direct cargo or co-trafficked with Rab11 endosomes unresolved
    • Quantitative contribution to surface TCR-CD3 levels not fully defined
  12. 2024 Medium

    Connected FIP3 recycling activity to immune evasion by showing it recycles PD-L1 to the gastric cancer-cell surface under IKZF4/NONO transcriptional control.

    Evidence FIP3 KD/OE, PD-L1 surface assays, T-cell cytotoxicity, in vivo tumor experiments, and transcription-factor knockout

    PMID:38211652

    Open questions at the time
    • Direct molecular link between FIP3 and PD-L1 cargo selection not defined
    • Single cancer-type model

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RAB11FIP3 integrates competing Rab11- and Arf6-dependent functions and selects specific cargo (Rac1, Lck, PD-L1) for synapse versus midbody delivery remains unresolved.
  • No structural model of the full FIP3 scaffold with simultaneous effector engagement
  • Cargo-selection mechanism for distinct trafficking routes unknown
  • Switch coordinating motor recruitment with phosphorylation state undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 3 GO:0008092 cytoskeletal protein binding 2
Localization
GO:0005768 endosome 3 GO:0005815 microtubule organizing center 2 GO:0005886 plasma membrane 2 GO:0005829 cytosol 1
Pathway
R-HSA-1640170 Cell Cycle 3 R-HSA-168256 Immune System 3 R-HSA-5653656 Vesicle-mediated transport 3 R-HSA-9609507 Protein localization 2
Partners
ARF6DYNC1LI1DYNC1LI2LCKRAB11ARAC1

Evidence

Reading pass · 12 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2001 Eferin (RAB11FIP3) was identified as a novel Rab11 effector protein. A 20-amino acid domain at the C-terminus of Eferin (and related Rab11/25-interacting proteins Rip11 and nRip11) was shown to be necessary and sufficient for Rab11 binding in vitro and required for correct localization of Rab11 effector proteins in vivo. Biochemical binding assays (in vitro Rab11 binding), mutagenesis of the C-terminal domain, in vivo localization studies The Journal of biological chemistry High 11481332
2004 RAB11FIP3 localizes to a Rab11-positive pericentrosomal compartment during interphase and to the cleavage furrow during cytokinesis. This localization is dependent on both microtubule and actin filament integrity. Expression of an N-terminally truncated mutant (Rab11-FIP3(244-756)) did not inhibit transferrin recycling, suggesting RAB11FIP3 is not required for transferrin recycling. Immunofluorescence microscopy with specific polyclonal antibody, cytoskeletal disruption experiments, dominant-negative mutant expression, transferrin recycling assay Biochemical and biophysical research communications Medium 15158446
2007 RAB11FIP3 is critical for the structural integrity of the endosomal recycling compartment (ERC). Knockdown of RAB11FIP3 or expression of a Rab11-binding-deficient mutant caused loss of all ERC-marker protein staining from the pericentrosomal region and blocked fluorophore-labelled transferrin from accessing the pericentrosomal region. A coiled-coil region between residues 463–692 that exists as a dimer in solution is critical for RAB11FIP3 function on the ERC. siRNA knockdown, Rab11-binding-deficient mutant expression, immunofluorescence microscopy, transferrin trafficking assay, biochemical analysis of coiled-coil domain dimerization Traffic (Copenhagen, Denmark) High 17394487
2007 RAB11FIP3 predominantly interacts with Arf6 (over Arf4/5) in vitro and in vivo. FIP3 binds to the Arf6 C-terminus rather than the switch motifs. Arf6 binding to FIP3 is required for targeting Arf6 to the cleavage furrow during cytokinesis, and FIP3 acts as a scaffolding protein for Arf6 recruitment to the midbody during late telophase. Biochemical affinity measurements (various Arf GTPases), co-immunoprecipitation, domain mapping/mutagenesis, live-cell imaging during cytokinesis European journal of cell biology High 17628206
2009 RAB11FIP3 directly interacts with dynein light intermediate chain 1 (DLIC-1, DYNC1LI1), a subunit of cytoplasmic dynein 1. Rab11a, FIP3, and DLIC-1 form a ternary complex. FIP3 recruits DLIC-1 onto membranes, and association between FIP3 and DLIC-1 at the cell periphery precedes minus-end-directed microtubule-based transport. Knockdown of DLIC-1 inhibits pericentrosomal accumulation of ERC proteins. A DLIC-1-binding truncation mutant of FIP3 disrupts ERC protein pericentrosomal accumulation. Co-immunoprecipitation, GST pulldown, immunofluorescence co-localization, siRNA knockdown, dominant-negative mutant expression Journal of cell science High 20026645
2009 RAB11FIP3 is associated with recycling endosomes at the leading edge of motile breast carcinoma cells. FIP3 is required for motility of MDA-MB-231 cells, regulates Rac1-dependent actin cytoskeleton dynamics and lamellipodia formation/ruffling, and controls Arf6 localization at the plasma membrane of migrating cells. siRNA knockdown, cell motility assays, immunofluorescence microscopy, actin dynamics assessment European journal of cell biology Medium 19327867
2010 RAB11FIP3 also binds dynein light intermediate chain 2 (DLIC-2, DYNC1LI2) via its N-terminal 435 amino acids and links Rab11a to DLIC-2. FIP3 recruits DLIC-2 onto membranes. DLIC-2 is necessary for pericentrosomal accumulation of endocytosed transferrin at the ERC. Overexpression of FIP3 fragments the Golgi complex by sequestering cytoplasmic dynein-1. Co-immunoprecipitation, membrane recruitment assay, siRNA knockdown of DLIC-2, transferrin trafficking assay, overexpression with Golgi morphology assessment Biochemical and biophysical research communications Medium 20214888
2011 FIP3 localization to recycling endosomes in interphase and their transport to the intercellular bridge during cytokinesis depend on Rab11. Targeting of FIP3-positive endosomal vesicles to the Flemming body (midbody) in the abscission phase depends on Arf6. After abscission, FIP3 and Arf6 are incorporated into one daughter cell as a Flemming body remnant. Detection of endogenous proteins by immunofluorescence, time-lapse analysis of FIP3, Rab11, and Arf6 localization during cytokinesis Genes to cells : devoted to molecular & cellular mechanisms Medium 21790911
2012 RAB11FIP3 is a cell cycle-regulated phosphoprotein with four in vivo phosphorylation sites (S-102, S-280, S-347, S-450). S-102 is a Cdk1-cyclin B substrate in vitro, is phosphorylated in metaphase, and dephosphorylated as cells enter telophase. Expression of FIP3-S102D increased binucleate cell frequency. Dephosphorylation of FIP3 accompanies its translocation from the cytosol to membranes during telophase, but the identified phospho-acceptor sites do not control spatial recycling endosome distribution. Mass spectrometry phosphosite identification, in vitro Cdk1-cyclin B kinase assay, phospho-specific analysis across cell cycle stages, phosphomimetic/phospho-dead mutant expression, binucleate cell frequency scoring BMC cell biology Medium 22401586
2016 RAB11FIP3 controls Rac1 intracellular localization and targeting to the immunological synapse. A significant fraction of Rac1 associates with Rab11-positive recycling endosomes, and FIP3 mediates this association. FIP3 regulates T-cell spreading, synapse symmetry, and T-cell activation leading to cytokine production in a Rac1-dependent manner. Overexpression and siRNA silencing of FIP3, co-localization studies (Rac1 with Rab11-positive endosomes), live-cell imaging, T-cell activation assays (cytokine production) The EMBO journal Medium 27154205
2017 RAB11FIP3-dependent endosomal trafficking of Lck controls its delivery to the immunological synapse and modulates early TCR signaling events including tyrosine phosphorylation of TCRζ, ZAP70, and LAT, intracellular calcium levels, and IL-2 gene expression. FIP3's interaction with Rab11 is required for proper Lck subcellular localization. FIP3 modulates TCR-CD3 cell surface expression via regulation of steady-state Lck-mediated TCRζ phosphorylation. FIP3 overexpression and siRNA silencing, Rab11-interaction mutants, immunofluorescence localization of Lck, phosphotyrosine western blotting, calcium flux assays, IL-2 reporter assays Journal of immunology (Baltimore, Md. : 1950) Medium 28235866
2024 RAB11FIP3 facilitates endosomal recycling of PD-L1 to the cell membrane of gastric cancer cells. The transcription factors IKZF4 and NONO synergistically upregulate RAB11FIP3 expression. Silencing of RAB11FIP3 reduces PD-L1 surface expression, promotes T-cell proliferation and cytotoxicity towards gastric cancer cells, and inhibits tumor immune evasion in vivo. RAB11FIP3 overexpression and siRNA silencing, PD-L1 cell-surface expression assay, T-cell cytotoxicity assays, in vivo mouse tumor experiments with CD8+ T-cell infiltration analysis, transcription factor KO experiments Cancer letters Medium 38211652

Source papers

Stage 0 corpus · 15 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2009 Rab11-FIP3 links the Rab11 GTPase and cytoplasmic dynein to mediate transport to the endosomal-recycling compartment. Journal of cell science 160 20026645
2001 Identification of a novel Rab11/25 binding domain present in Eferin and Rip proteins. The Journal of biological chemistry 94 11481332
2004 Rab11-FIP3 localises to a Rab11-positive pericentrosomal compartment during interphase and to the cleavage furrow during cytokinesis. Biochemical and biophysical research communications 87 15158446
2007 Rab11-FIP3 is critical for the structural integrity of the endosomal recycling compartment. Traffic (Copenhagen, Denmark) 61 17394487
2010 Rab11-FIP3 binds dynein light intermediate chain 2 and its overexpression fragments the Golgi complex. Biochemical and biophysical research communications 56 20214888
2016 Rac1-Rab11-FIP3 regulatory hub coordinates vesicle traffic with actin remodeling and T-cell activation. The EMBO journal 47 27154205
2007 Molecular characterization of Rab11-FIP3 binding to ARF GTPases. European journal of cell biology 44 17628206
2009 Rab11-FIP3 is a Rab11-binding protein that regulates breast cancer cell motility by modulating the actin cytoskeleton. European journal of cell biology 41 19327867
2011 Distinct roles of Rab11 and Arf6 in the regulation of Rab11-FIP3/arfophilin-1 localization in mitotic cells. Genes to cells : devoted to molecular & cellular mechanisms 33 21790911
2017 Increased Eps15 homology domain 1 and RAB11FIP3 expression regulate breast cancer progression via promoting epithelial growth factor receptor recycling. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine 24 28215104
2017 Rab11-FIP3 Regulation of Lck Endosomal Traffic Controls TCR Signal Transduction. Journal of immunology (Baltimore, Md. : 1950) 24 28235866
2005 Purification and functional properties of Rab11-FIP3. Methods in enzymology 18 16473615
2024 IKZF4/NONO-RAB11FIP3 axis promotes immune evasion in gastric cancer via facilitating PD-L1 endosome recycling. Cancer letters 17 38211652
2016 The functional interplay of Rab11, FIP3 and Rho proteins on the endosomal recycling pathway controls cell shape and symmetry. Small GTPases 13 27533792
2012 Rab11-FIP3 is a cell cycle-regulated phosphoprotein. BMC cell biology 12 22401586

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