Affinage

RASAL3

RAS protein activator like-3 · UniProt Q86YV0

Length
1011 aa
Mass
111.9 kDa
Annotated
2026-06-10
23 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

RASAL3 is a hematopoietically enriched RasGAP that negatively regulates Ras/ERK signaling to control immune cell survival, expansion, and inflammatory output (PMID:25793935, PMID:25652366). Biochemically it possesses RasGAP activity without Rap1GAP activity and represses TCR- and α-GalCer-stimulated ERK phosphorylation (PMID:25793935, PMID:25652366); its catalytic domain also stimulates GTP hydrolysis of the Rho-family GTPase Rac2 in vitro (PMID:30271600). Loss of RASAL3 reduces survival of naive and activated T cells through increased apoptosis and lowered Bcl2, depletes hepatic NKT cells, and attenuates contact-hypersensitivity and α-GalCer-induced liver damage, establishing RASAL3 as a survival factor that sets the magnitude of T/NKT cell responses (PMID:25793935, PMID:25652366, PMID:29291408). In neutrophils RASAL3 is inducible by inflammatory stimuli and restrains neutrophil hyperinflammation, with its deficiency driving lethal septic organ damage (PMID:34777356). In migrating neutrophils and macrophages, spatially restricted RASAL3 activity locally suppresses Ras to extinguish protrusions at the cell front, while rear or uniform recruitment paradoxically enhances polarity and speed via myosin II-dependent rear contraction and mTORC2-dependent front F-actin assembly, defining a Ras-controlled front/back feedback loop independent of PI3K (PMID:37220748, PMID:38951708). RASAL3 physically associates with CCDC88B and the Rho/Rac GEF ARHGEF2 to oppositely tune RHOA activation during dendritic cell migration (PMID:38200184). Beyond immunity, RASAL3 is epigenetically silenced in prostate cancer-associated fibroblasts to derepress oncogenic Ras-driven macropinocytosis and glutamine secretion that fuels tumor progression (PMID:30047926), it is bound by CD229/SLAMF3 to activate RAS/ERK in multiple myeloma (PMID:36445333), and its mRNA stability is controlled by the m6A demethylase ALKBH5 (PMID:36876119).

Mechanistic history

Synthesis pass · year-by-year structured walk · 11 steps
  1. 2015 High

    Established RASAL3 as a bona fide RasGAP whose loss compromises peripheral T cell survival, defining its core role as a negative regulator of Ras/ERK that maintains the naive T cell pool.

    Evidence In vitro RasGAP/Rap1GAP assays plus systemic knockout mice with adoptive transfer and apoptosis readouts

    PMID:25793935

    Open questions at the time
    • Mechanism linking ERK suppression to survival/apoptosis not resolved
    • Thymic development unaffected, leaving the developmental context of RASAL3 unexplained
  2. 2015 High

    Extended RASAL3's Ras-suppressive role to NKT cells, showing it limits NKT expansion and effector cytokine output and that its loss dysregulates antigen-driven ERK activation.

    Evidence Systemic knockout mice with α-GalCer stimulation, ERK phosphorylation and cytokine assays

    PMID:25652366

    Open questions at the time
    • Whether NKT depletion reflects survival vs developmental defect not fully separated
    • Direct Ras-GTP measurement in NKT cells not shown
  3. 2017 Medium

    Showed RASAL3 is also required for survival of activated T cells, linking it to control of inflammatory response magnitude in vivo.

    Evidence Knockout mice in Th1/Th2 contact hypersensitivity models with Bcl2 analysis

    PMID:29291408

    Open questions at the time
    • Single lab, single study
    • Causal link between Bcl2 loss and apoptosis not mechanistically dissected
  4. 2018 Medium

    Revealed a non-immune oncogenic axis: epigenetic silencing of RASAL3 in cancer-associated fibroblasts derepresses Ras to drive macropinocytic glutamine supply to prostate tumor cells.

    Evidence Orthotopic xenografts with macropinocytosis/glutamine inhibition and methylation analysis

    PMID:30047926

    Open questions at the time
    • Direct demonstration that RASAL3 re-expression reverses the metabolic phenotype not detailed
    • Single lab
  5. 2018 Medium

    Demonstrated the RASAL3 catalytic domain can act on Rac2 in addition to Ras, expanding its substrate repertoire to a Rho-family GTPase.

    Evidence In vitro GTPase assay with recombinant RASAL3 catalytic domain and Rac2

    PMID:30271600

    Open questions at the time
    • No catalytic-dead mutagenesis to confirm specificity
    • Cellular relevance of Rac2 activity not established
  6. 2021 Medium

    Identified RASAL3 as an inflammation-inducible brake on neutrophil responses whose loss produces lethal hyperinflammation, broadening its function to innate immunity.

    Evidence Knockout mice in septic shock and sickle cell disease models with neutrophil functional assays

    PMID:34777356

    Open questions at the time
    • Molecular trigger of inflammation-induced RASAL3 upregulation unknown
    • Single lab
  7. 2023 High

    Used optogenetics to prove RASAL3 acts as a spatially deployable Ras suppressor that directly governs protrusion, cell shape, and migration mode via mTORC2 rather than PI3K.

    Evidence Optogenetic recruitment in HL-60 neutrophils and RAW 264.7 macrophages with mTORC2/PI3K inhibitor epistasis (preprint and Developmental Cell)

    PMID:37220748 PMID:37693515

    Open questions at the time
    • Endogenous spatial regulators of RASAL3 localization not identified
    • Connection to in vivo immune migration not directly tested
  8. 2024 High

    Resolved the front/back logic of RASAL3 action, showing rear Ras suppression drives polarity through myosin II contraction coupled to mTORC2-dependent front actin assembly in a feedback loop.

    Evidence Optogenetics with blebbistatin and mTORC2 inhibition plus computational modeling across two cell types

    PMID:38951708

    Open questions at the time
    • How endogenous signaling sets RASAL3 polarity in vivo unknown
    • Quantitative link between Ras-GTP gradients and contractility not fully defined
  9. 2024 Medium

    Placed RASAL3 in a physical complex with CCDC88B and ARHGEF2 that tunes RHOA-dependent dendritic cell migration, connecting it to a Rho-GEF regulatory module.

    Evidence Co-immunoprecipitation, Rasal3-mutant mice, DC migration and RHOA activity assays in neuroinflammation/colitis models

    PMID:38200184

    Open questions at the time
    • Direct vs indirect nature of each interaction not fully mapped
    • How the complex spatially integrates RHOA vs Ras regulation unresolved
  10. 2022 Medium

    Identified CD229/SLAMF3 as a RASAL3-binding receptor that channels into RAS/ERK activation to promote myeloma proliferation.

    Evidence Co-IP/mass spectrometry, co-culture immunofluorescence, proliferation assays and xenografts

    PMID:36445333

    Open questions at the time
    • Whether CD229 binding inhibits RASAL3 GAP activity not biochemically shown
    • Single lab
  11. 2023 Medium

    Showed RASAL3 abundance is set post-transcriptionally by ALKBH5-mediated m6A regulation of its mRNA, coupling Ras pathway output to RNA modification in cardiac stress.

    Evidence Alkbh5 genetic mouse models, m6A-seq, mRNA stability assays and RAS/ERK analysis

    PMID:36876119

    Open questions at the time
    • Direct m6A sites on Rasal3 mRNA and reader identity not fully defined
    • Single lab

Open questions

Synthesis pass · forward-looking unresolved questions
  • How endogenous upstream signals spatially position and activate RASAL3, and whether its dual Ras/Rac2 GAP activities are differentially deployed across cell types, remains unresolved.
  • No structure of RASAL3 or its regulatory domains in the corpus
  • Physiological balance between RasGAP and Rac2-GAP activity unknown
  • Endogenous recruitment cues for migratory polarity unidentified

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0098772 molecular function regulator activity 3 GO:0140096 catalytic activity, acting on a protein 2
Localization
GO:0005886 plasma membrane 2
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-168256 Immune System 4
Complex memberships
CCDC88B/RASAL3/ARHGEF2 complex

Evidence

Reading pass · 12 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2015 RASAL3 possesses RasGAP activity but not Rap1GAP activity, and represses TCR-stimulated ERK phosphorylation in T cells. In systemic Rasal3-deficient mice, naive CD4 and CD8 T cells in the periphery were significantly reduced due to increased apoptosis, while thymic T cell development (positive, negative, and β-selection) was unaffected. In vivo adoptive transfer showed impaired survival of Rasal3-deficient naive CD4 T cells, whereas IL-7-dependent survival in vitro was unaltered. In vitro RasGAP/Rap1GAP activity assays, systemic Rasal3-knockout mice, flow cytometry, adoptive transfer, apoptosis assays PloS one High 25793935
2015 RASAL3 is predominantly expressed in hematopoietic lineages (NKT, B, and T cells) and negatively regulates Ras/ERK signaling in NKT cells. RASAL3-deficient mice showed severe decrease in NKT cells in the liver, and RASAL3-deficient NKT cells treated with α-GalCer exhibited augmented ERK phosphorylation, confirming dysregulated Ras signaling. Loss of RASAL3 attenuated α-GalCer-induced liver damage and reduced IL-4 and IFN-γ production from NKT cells. Systemic RASAL3-knockout mice, α-GalCer stimulation, flow cytometry, ERK phosphorylation assays, cytokine measurement European journal of immunology High 25652366
2017 Rasal3-deficient mice show ameliorated Th1- and Th2-dependent contact hypersensitivity reactions and a shortage of T cells at regional lymph nodes. Activated Rasal3-deficient T cells showed increased cell death with reduced Bcl2 expression, indicating that Rasal3 is required for survival of both naive and activated T cells to control the magnitude of inflammatory responses. Systemic Rasal3-knockout mice, contact hypersensitivity models, flow cytometry, Bcl2 expression analysis, in vivo T cell transfer Biochemical and biophysical research communications Medium 29291408
2018 RASAL3 is epigenetically silenced (promoter hypermethylation) in prostatic cancer-associated fibroblasts (CAF), leading to oncogenic Ras activation. This Ras activity drives macropinocytosis-mediated glutamine synthesis in CAF, providing glutamine to prostate cancer epithelial cells to fuel proliferation and promote neuroendocrine differentiation. ADT further promotes RASAL3 epigenetic silencing and glutamine secretion. Orthotopic xenograft models, macropinocytosis inhibition, glutamine transport inhibition, epigenetic analysis (methylation), castration-resistant xenograft rescue experiments The Journal of clinical investigation Medium 30047926
2018 The catalytic (GAP) domain of RASAL3 interacts with and stimulates GTP hydrolysis of the Rho family GTPase Rac2 in vitro, in addition to its known activity on p21ras. By contrast, p50 rhoGAP did not markedly affect Rac2 GTPase activity but did accelerate activity of Rac1, RhoA, and Cdc42. In vitro GTPase activity assay with recombinant RASAL3 catalytic domain and Rac2 Biomedical reports Medium 30271600
2021 RASAL3 is highly expressed in neutrophils and its expression is upregulated by exogenous inflammatory stimuli. RASAL3 deficiency (KO mice) triggers augmented neutrophil responses and hyperinflammation in acute inflammatory conditions, leading to accelerated mortality in a septic shock model via severe organ damage. Mice modeling sickle cell disease were found to have low neutrophil RASAL3 expression upon LPS activation, and this correlated with excessive neutrophilic hyperinflammation and increased mortality. RASAL3-KO mice, septic shock model, sickle cell disease mouse model, LPS stimulation, neutrophil functional assays, histology Frontiers in immunology Medium 34777356
2023 Optogenetic recruitment of RASAL3 to the cell front of differentiated HL-60 neutrophils or RAW 264.7 macrophages extinguished protrusions and reversed migration direction. Global or rear-targeted RASAL3 recruitment caused cells to polarize and move more rapidly, effects that were mTORC2-dependent but relatively independent of PI3K. These findings demonstrate that local Ras suppression by RASAL3 directly controls actin assembly, cell shape, and migration mode in immune cells. Optogenetics (iLID/SspB system), HL-60 neutrophils and RAW 264.7 macrophages, live-cell imaging, pharmacological inhibition of mTORC2 and PI3K Developmental cell High 37220748
2024 Optogenetic recruitment of RASAL3 to the front of HL-60 neutrophils or macrophages extinguished protrusions and changed migration direction. Uniform or rear-targeted RASAL3 recruitment paradoxically increased cell polarization and migration speed through myosin II-dependent rear contraction followed by mTORC2-dependent actin polymerization at the front. RASAL3 thus controls a front/back feedback loop in which Ras suppression at the rear drives enhanced polarity and motility. Optogenetics, HL-60 neutrophils, RAW 264.7 macrophages, myosin II inhibition (blebbistatin), mTORC2 inhibition, computational modeling, live-cell imaging Nature cell biology High 38951708
2024 RASAL3 physically interacts with CCDC88B and ARHGEF2 (a Rho/Rac GEF) as identified by co-immunoprecipitation. The CCDC88B/RASAL3/ARHGEF2 complex regulates dendritic cell migration by modulating RHOA activation, with ARHGEF2 and RASAL3 acting in opposing regulatory fashions. Rasal3-mutant mice show dampened neuroinflammation and altered susceptibility to colitis, and Rasal3-mutant DCs show enhanced migratory properties in vitro. Co-immunoprecipitation, Rasal3-mutant mice, DC migration assays, neuroinflammation and colitis disease models, RHOA activity assays Communications biology Medium 38200184
2022 CD229 (SLAMF3) interacts with RASAL3 as identified by co-immunoprecipitation coupled with mass spectrometry. Intercellular tyrosine phosphorylation-mediated self-activation of CD229 activates the RAS/ERK signaling pathway via interaction with RASAL3 to promote multiple myeloma cell proliferation. Co-immunoprecipitation coupled with mass spectrometry, immunofluorescence co-culture assay, xenograft mouse model, in vitro proliferation assays Aging Medium 36445333
2023 ALKBH5 (m6A demethylase) regulates Rasal3 mRNA stability in an m6A-dependent manner through posttranscriptional regulation. ALKBH5-mediated reduction of Rasal3 mRNA stability activates RAS signaling and inhibits apoptosis through the RAS/RAF/ERK pathway, thereby alleviating doxorubicin-induced cardiotoxicity. Alkbh5 whole-body KO and myocardial-specific KO mice showed increased mortality and aggravated cardiac injury, while ALKBH5 overexpression was protective. Alkbh5-knockout, knockin, and myocardial-specific KO mice, m6A-sequencing, mRNA stability assays, cardiac function assessment, RAS/ERK signaling analysis iScience Medium 36876119
2023 Optogenetic targeting of RASAL3 to the cell front of HL-60 neutrophils extinguished existing protrusions and changed migration direction. Global or rear-targeted RASAL3 caused rapid cell polarization and faster migration, mediated by increased actomyosin contractility at the back and sustained localized F-actin polymerization at the front, with effects captured by computational simulations of Ras-controlled front/back feedback loops. Optogenetics, HL-60 neutrophils, Dictyostelium (C2GAPB ortholog comparison), computational modeling, live-cell imaging bioRxivpreprint Medium 37693515

Source papers

Stage 0 corpus · 23 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2018 Stromal epigenetic alterations drive metabolic and neuroendocrine prostate cancer reprogramming. The Journal of clinical investigation 142 30047926
2019 Epigenetic changes in fibroblasts drive cancer metabolism and differentiation. Endocrine-related cancer 37 31627186
2018 Transcriptome Analysis of Bronchoalveolar Lavage Fluid From Children With Mycoplasma pneumoniae Pneumonia Reveals Natural Killer and T Cell-Proliferation Responses. Frontiers in immunology 36 29967623
2015 RASAL3, a novel hematopoietic RasGAP protein, regulates the number and functions of NKT cells. European journal of immunology 32 25652366
2023 Actuation of single downstream nodes in growth factor network steers immune cell migration. Developmental cell 31 37220748
2015 The Ras GTPase-activating protein Rasal3 supports survival of naive T cells. PloS one 21 25793935
2024 Ras suppression potentiates rear actomyosin contractility-driven cell polarization and migration. Nature cell biology 20 38951708
2017 RNA-seq implicates deregulation of the immune system in the pathogenesis of diverticulitis. American journal of physiology. Gastrointestinal and liver physiology 19 28619727
2015 Identification of potential mutations and genomic alterations in the epithelial and spindle cell components of biphasic synovial sarcomas using a human exome SNP chip. BMC medical genomics 19 26503545
2021 Differential DNA Methylation Profiles in Patients with Temporal Lobe Epilepsy and Hippocampal Sclerosis ILAE Type I. Journal of molecular neuroscience : MN 18 33403589
2020 Digenic inheritance of subclinical variants in Noonan Syndrome patients: an alternative pathogenic model? European journal of human genetics : EJHG 17 32514133
2021 RASAL3 Is a Putative RasGAP Modulating Inflammatory Response by Neutrophils. Frontiers in immunology 16 34777356
2021 Transcriptomics unravels molecular players shaping dorsal lip hypertrophy in the vacuum cleaner cichlid, Gnathochromis permaxillaris. BMC genomics 13 34225643
2017 Rasal3-mediated T cell survival is essential for inflammatory responses. Biochemical and biophysical research communications 13 29291408
2023 m6A demethylase ALKBH5 attenuates doxorubicin-induced cardiotoxicity via posttranscriptional stabilization of Rasal3. iScience 9 36876119
2020 The impact of blood-processing time on the proteome of human peripheral blood mononuclear cells. Biochimica et biophysica acta. Proteins and proteomics 9 33301959
2024 CCDC88B interacts with RASAL3 and ARHGEF2 and regulates dendritic cell function in neuroinflammation and colitis. Communications biology 7 38200184
2021 DNA Methylome Mapping Identifies Epigenetic Abnormalities in Intestinal Lymphocyte Regulation in Human Necrotizing Enterocolitis. Digestive diseases and sciences 7 34846677
2021 [Expression of RASGRP2 in Lung Adenocarcinoma and Its Effect 
on Immune Microenvironment]. Zhongguo fei ai za zhi = Chinese journal of lung cancer 5 34157800
2022 CD229 interacts with RASAL3 to activate RAS/ERK pathway in multiple myeloma proliferation. Aging 4 36445333
2019 Change of Ras and its guanosine triphosphatases (GTPases) during development and regression in bovine corpus luteum. Theriogenology 3 31887652
2023 Ras-mediated homeostatic control of front-back signaling dictates cell polarity. bioRxiv : the preprint server for biology 1 37693515
2018 RASAL3 preferentially stimulates GTP hydrolysis of the Rho family small GTPase Rac2. Biomedical reports 1 30271600

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