Affinage

RGL3

Ral guanine nucleotide dissociation stimulator-like 3 · UniProt Q3MIN7

Length
710 aa
Mass
78.1 kDa
Annotated
2026-06-10
29 papers in source corpus 7 papers cited in narrative 7 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 5/5 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

RGL3 is a RalGEF-family guanine nucleotide exchange factor that couples activated Ras-superfamily GTPases to Ral activation, functioning as a Ras effector in growth control (PMID:10869344, PMID:20801877). Through a C-terminal Ras-binding domain it engages GTP-loaded Rit and Ras in a GTP- and effector-loop-dependent manner and catalyzes guanine nucleotide exchange on Ral in vivo when Rit or Ras is activated (PMID:10869344); its binding repertoire extends to M-Ras, H-Ras, N-Ras, Rap1, Rap2, and R-Ras, but not Ral or Rho (PMID:11313946, PMID:17382517). RGL3 localizes predominantly to the cytoplasm, and plasma-membrane targeting that mimics persistent Ras activation enhances its growth-transforming activity, while silencing it impairs anchorage-dependent and anchorage-independent growth of non-small cell lung cancer cells (PMID:17382517, PMID:20801877). An N-terminal proline cluster recruits profilin II, and the two cooperate to enhance N-Ras-induced focus formation, linking RGL3 to actin-dependent control of cell spreading and morphology (PMID:17382517). Distinct from its transforming role, RGL3 inhibits Elk-1-dependent transcription downstream of activated H-Ras or MEKK-1 through a Ras-binding-domain-independent mechanism (PMID:11313946).

Mechanistic history

Synthesis pass · year-by-year structured walk · 7 steps
  1. 2000 Medium

    Established RGL3 as a bona fide RalGEF by showing it binds activated Rit and Ras through a defined C-terminal domain and stimulates Ral nucleotide exchange in vivo, placing it downstream of these GTPases.

    Evidence Yeast two-hybrid for Rit-binding partners with GTP- and effector-loop-dependence tests and an in vivo RalGEF activity assay

    PMID:10869344

    Open questions at the time
    • No direct biochemical reconstitution of exchange activity with purified components
    • Endogenous RGL3-Ral coupling not demonstrated
  2. 2001 Medium

    Revealed a paradoxical signaling role: rather than activating Elk-1 like other RalGEFs, RGL3 binds GTP-bound M-Ras and Ras yet inhibits Ras/MEKK-1-driven Elk-1 transcription via a Ras-binding-domain-independent mechanism, and suppresses Src-transformed fibroblast growth.

    Evidence Yeast two-hybrid and co-immunoprecipitation binding, Elk-1 reporter assays, domain deletion analysis, and transformed cell growth assays

    PMID:11313946

    Open questions at the time
    • The RBD-independent inhibitory mechanism and the required second signal are unidentified
    • Relationship between transcriptional inhibition and Ral activation unresolved
  3. 2007 Medium

    Broadened the GTPase interaction map and identified profilin II as an N-terminal proline-cluster partner, connecting RGL3 to actin-dependent morphology while showing opposing effects on cell spreading versus oncogenic focus formation.

    Evidence Yeast two-hybrid, cytoplasmic localization in fibroblasts, cell spreading and focus-formation assays, and proline-cluster mutation mapping

    PMID:17382517

    Open questions at the time
    • Mechanism linking profilin II recruitment to focus formation undefined
    • How RGL3 suppresses spreading yet enhances transformation not reconciled
  4. 2010 Medium

    Demonstrated RGL3 functions in the Ras effector RalGEF family to support transformation, with membrane targeting enhancing its transforming activity and silencing impairing lung cancer cell growth.

    Evidence Plasma membrane-targeting constructs, soft-agar growth assays, and siRNA knockdown in NSCLC lines

    PMID:20801877

    Open questions at the time
    • Whether the growth phenotype depends on Ral activation per se untested
    • No in vivo tumor model
  5. 2016 Low

    Reinforced a functional requirement for RGL3 in lung cancer cell proliferation under both anchorage conditions.

    Evidence siRNA knockdown with anchorage-dependent and -independent growth assays in NSCLC lines

    PMID:27149377

    Open questions at the time
    • Single knockdown approach without rescue
    • Downstream effectors mediating the phenotype not identified
  6. 2020 Low

    Resolved the RIT1-RGL3 interaction interface at high resolution, refining the structural basis of the GTPase-effector contact.

    Evidence DoMY-Seq (yeast two-hybrid coupled to next-generation sequencing) fragment-library domain mapping

    PMID:33410398

    Open questions at the time
    • Yeast two-hybrid-based mapping without structural or biochemical confirmation
    • Functional consequence of the mapped interface untested
  7. 2006 Low

    Placed RGL3 within signal-responsive phosphoregulation by detecting vasopressin-dependent changes in its phosphorylation state.

    Evidence IMAC phosphopeptide enrichment with LC-MS/MS and label-free quantification in rat inner medullary collecting duct cells

    PMID:16641100

    Open questions at the time
    • Phosphosites and responsible kinase not defined
    • Functional effect of phosphorylation on RGL3 activity unknown

Open questions

Synthesis pass · forward-looking unresolved questions
  • How RGL3's RalGEF activity, its RBD-independent inhibition of Elk-1 transcription, and its profilin II-actin axis integrate into a single coherent function in normal and transformed cells remains unresolved.
  • No structural model of full-length RGL3
  • Endogenous substrate and effector hierarchy not established
  • Physiological context outside cancer cell lines undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060089 molecular transducer activity 3 GO:0098772 molecular function regulator activity 2 GO:0140096 catalytic activity, acting on a protein 1
Localization
GO:0005829 cytosol 1
Pathway
R-HSA-162582 Signal Transduction 3
Partners
HRASMRASNRASPFN2RAP1RIT1RRAS

Evidence

Reading pass · 7 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2000 RGL3 was identified as a novel RalGEF-family protein that interacts with activated (GTP-bound) Rit and Ras in a GTP- and effector loop-dependent manner via a C-terminal 99-amino acid domain, and exhibits guanine nucleotide exchange activity toward the small GTPase Ral that is stimulated in vivo by expression of either activated Rit or Ras. Yeast two-hybrid screening for Rit-binding proteins; in vivo RalGEF activity assay; GTP-dependence and effector loop-dependence tests The Journal of biological chemistry Medium 10869344
2001 RPM/RGL3 interacts selectively and strongly with GTP-bound M-Ras and p21 Ras; unlike other RalGEFs, transient expression of RPM/RGL3 did not activate an Elk-1 reporter gene but strongly inhibited Elk-1-dependent gene induction triggered by activated H-Ras or MEKK-1. This inhibitory effect was independent of the Ras binding domain and required a second signal from p21 Ras or MEKK-1 (but not Raf-1 or M-Ras). RPM/RGL3 also strongly inhibited growth of Src-Y527F-transformed fibroblasts. Yeast two-hybrid and co-immunoprecipitation for binding; Elk-1 reporter gene assay; domain deletion analysis; cell growth/transformation assay Oncogene Medium 11313946
2007 Rgl3 was identified as a binding partner for Rap1, Rap2, H-Ras, N-Ras, and R-Ras (but not Ral or Rho) via yeast two-hybrid; Rgl3 localizes predominantly to the cytoplasm in fibroblasts; Rgl3 suppresses cell spreading induced by Rap1, R-Ras, or C3G-CAAX in HEK-293 cells; Rgl3 enhances focus formation by oncogenic H-Ras and N-Ras in NIH3T3 cells. Additionally, profilin II was identified as a binding partner for Rgl3 through its N-terminal proline cluster, and profilin II and Rgl3 cooperated to enhance N-Ras-induced focus formation. Yeast two-hybrid screening; subcellular localization by expression in fibroblasts; cell spreading assay; focus-formation assay; domain mapping by proline cluster mutation Cellular signalling Medium 17382517
2010 RGL3, along with RalGDS, Rgl1, and Rgl2, functions as a Ras effector RalGEF; plasma membrane targeting (mimicking persistent Ras activation) enhanced the growth-transforming activity of RGL3. Silencing of RGL3 in non-small cell lung cancer cell lines inhibited anchorage-dependent and anchorage-independent growth. Plasma membrane-targeting constructs; soft-agar growth assay; siRNA knockdown; cell growth assays The Journal of biological chemistry Medium 20801877
2016 RGL3 silencing in non-small cell lung cancer (NSCLC) cell lines inhibited cell population growth under anchorage-dependent and anchorage-independent conditions, demonstrating a functional role in supporting lung cancer cell proliferation. siRNA knockdown; anchorage-dependent and -independent growth assays PloS one Low 27149377
2020 High-resolution domain mapping (DoMY-Seq) confirmed the RIT1-RGL3 protein-protein interaction and mapped the interacting interface at high resolution, validating the interaction between RIT1 and RGL3. DoMY-Seq (yeast two-hybrid coupled with next-generation sequencing); domain mapping via fragment library The Journal of biological chemistry Low 33410398
2006 Rgl3 was identified as a phosphoprotein whose phosphorylation state significantly changed in response to short-term vasopressin treatment in rat inner medullary collecting duct cells, as detected by label-free quantitative phosphoproteomics. Phosphopeptide enrichment by IMAC; LC-MS/MS neutral loss scanning; label-free quantification Proceedings of the National Academy of Sciences of the United States of America Low 16641100

Source papers

Stage 0 corpus · 29 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2006 Quantitative phosphoproteomics of vasopressin-sensitive renal cells: regulation of aquaporin-2 phosphorylation at two sites. Proceedings of the National Academy of Sciences of the United States of America 292 16641100
2023 Polygenic prediction of preeclampsia and gestational hypertension. Nature medicine 105 37248299
2008 Elucidating the germination transcriptional program using small molecules. Plant physiology 90 18359847
2010 RalGDS family members couple Ras to Ral signalling and that's not all. Cellular signalling 66 20478380
2017 GA-DELLA pathway is involved in regulation of nitrogen deficiency-induced anthocyanin accumulation. Plant cell reports 65 28275852
2000 A novel RalGEF-like protein, RGL3, as a candidate effector for rit and Ras. The Journal of biological chemistry 52 10869344
2010 Aberrant overexpression of the Rgl2 Ral small GTPase-specific guanine nucleotide exchange factor promotes pancreatic cancer growth through Ral-dependent and Ral-independent mechanisms. The Journal of biological chemistry 51 20801877
2011 GsGASA1 mediated root growth inhibition in response to chronic cold stress is marked by the accumulation of DELLAs. Journal of plant physiology 48 21855169
2013 Rit subfamily small GTPases: regulators in neuronal differentiation and survival. Cellular signalling 44 23770287
2024 Protein-altering variants at copy number-variable regions influence diverse human phenotypes. Nature genetics 36 38548989
2001 A novel potential effector of M-Ras and p21 Ras negatively regulates p21 Ras-mediated gene induction and cell growth. Oncogene 32 11313946
2021 The gibberellin signaling negative regulator RGA-LIKE3 promotes seed storage protein accumulation. Plant physiology 29 33793917
2013 The DELLA protein RGL3 positively contributes to jasmonate/ethylene defense responses. Plant signaling & behavior 22 23425858
2021 Multi-ancestry genome-wide gene-sleep interactions identify novel loci for blood pressure. Molecular psychiatry 21 33859359
2016 RalGPS2 Is Essential for Survival and Cell Cycle Progression of Lung Cancer Cells Independently of Its Established Substrates Ral GTPases. PloS one 17 27149377
2022 Polymorphisms of hypertension susceptibility genes as a risk factors of preeclampsia in the Caucasian population of central Russia. Placenta 16 36219912
2022 Aggregation of Genome-Wide Association Data from FinnGen and UK Biobank Replicates Multiple Risk Loci for Pregnancy Complications. Genes 15 36553520
2007 Identification of Rgl3 as a potential binding partner for Rap-family small G-proteins and profilin II. Cellular signalling 15 17382517
2023 Risk Effects of rs1799945 Polymorphism of the HFE Gene and Intergenic Interactions of GWAS-Significant Loci for Arterial Hypertension in the Caucasian Population of Central Russia. International journal of molecular sciences 13 37176017
2020 DoMY-Seq: A yeast two-hybrid-based technique for precision mapping of protein-protein interaction motifs. The Journal of biological chemistry 8 33410398
2024 Ratiometric gibberellin biosensors for the analysis of signaling dynamics and metabolism in plant protoplasts. The Plant journal : for cell and molecular biology 6 38525669
2024 Enhanced resolution profiling in twins reveals differential methylation signatures of type 2 diabetes with links to its complications. EBioMedicine 3 38574408
2023 Hidden protein-altering variants influence diverse human phenotypes. bioRxiv : the preprint server for biology 3 37333244
2026 GA2ox1-1 modulates bioactive gibberellin levels to regulate the RGL3-MYB169 pathway in pear stone cell formation. Plant physiology 2 41400294
2023 Whole Exome Sequencing Reveals Novel Candidate Genes in Familial Forms of Glaucomatous Neurodegeneration. Genes 2 36833422
2026 Precision nutrition for hypertension: tea, coffee, antioxidant vitamins interactions with polygenic risk in multi-ethnic populations. European journal of clinical nutrition 1 41501475
2024 Morphological classification of radial glia-like cells in the postnatal mouse subventricular zone. The European journal of neuroscience 1 39126378
2026 Distinct radial glia subtypes regulate midbrain dopaminergic neuron development. Nature neuroscience 0 41699318
2026 ARF6 integrates auxin and gibberellin signaling to promote stone cell lignification in pear via the HB49-MYB169 module. The New phytologist 0 42037589

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