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Showing NLE1 — RSA4 is a alias.

NLE1

Notchless protein homolog 1 · UniProt Q9NVX2

Length
485 aa
Mass
53.3 kDa
Annotated
2026-06-10
51 papers in source corpus 6 papers cited in narrative 8 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

NLE1 (Notchless Homolog 1) is a WD40-repeat protein that limits de novo protein biosynthesis and supports tumor cell proliferation, survival, and metastasis across multiple cancers (PMID:36219392). In colorectal cancer, NLE1 is transcriptionally controlled at an E-box in its promoter: c-MYC binds this element and prevents TGFβ-mediated downregulation, so SMAD4 loss leads to NLE1 upregulation (PMID:36219392). NLE1 is rate-limiting for protein synthesis, and its ablation activates p38/MAPK signaling, impairs autophagy with accumulation of p62- and LC3-positive structures, raises reactive oxygen species, and blocks proliferation, migration, invasion, and anchorage-independent growth (PMID:36219392). In vivo, NLE1 loss impairs orthotopic colorectal tumor growth and liver metastasis, and co-deletion of TP53 sensitizes NLE1-deficient cells to apoptosis (PMID:36219392). Beyond its biosynthetic role, NLE1 modulates Notch signaling: it forms a direct complex with Notch1 in brain tumor stem cells, where the NLE1-targeting drug edaravone impairs Notch signaling and sensitizes cells to radiation (PMID:39423824). In NSCLC and melanoma, NLE1 drives tumor phenotypes through PI3K/AKT signaling, and in NSCLC it activates E2F1-mediated transcription of CDK1, with CDK1 overexpression or Akt activation rescuing the effects of NLE1 knockdown (PMID:36818671, PMID:34019907).

Mechanistic history

Synthesis pass · year-by-year structured walk · 6 steps
  1. 2017 Medium

    Initial in vivo evidence placed NLE1 within the Notch1 signaling axis, linking its expression to cortical development and a neurological phenotype.

    Evidence Immunohistochemistry/qRT-PCR for NLE1 and Notch1 plus in vivo EEG with Notch agonist/antagonist in an absence epilepsy rat model

    PMID:28210849

    Open questions at the time
    • Correlative association between NLE1 and Notch1 without direct physical or biochemical interaction shown
    • No molecular mechanism connecting NLE1 to spike-and-wave discharges
    • Causality of NLE1 reduction not tested by gene manipulation
  2. 2021 Medium

    Loss-of-function in melanoma established NLE1 as a pro-tumorigenic factor acting through PI3K/AKT signaling, extending its role beyond Notch.

    Evidence shRNA knockdown in melanoma cell lines with proliferation/apoptosis/migration assays, xenografts, and PI3K/AKT western blotting

    PMID:34019907

    Open questions at the time
    • Mechanism by which NLE1 activates PI3K/AKT not defined
    • No direct binding partner identified
    • Single lab, single cancer type
  3. 2022 High

    A comprehensive colorectal cancer study defined the transcriptional control of NLE1 and its function as a limiting factor for protein biosynthesis driving tumor growth and metastasis.

    Evidence ChIP for c-MYC at the NLE1 promoter E-box, organoid and CRISPR-KO models, protein synthesis/autophagy/ROS assays, and an orthotopic metastasis model with TP53 epistasis

    PMID:36219392

    Open questions at the time
    • Direct molecular substrate of NLE1 in the protein biosynthesis machinery not biochemically defined
    • Mechanism linking NLE1 loss to p38/MAPK activation and autophagy impairment unresolved
    • Whether the biosynthetic role and Notch role are mechanistically connected is unknown
  4. 2023 Medium

    NSCLC studies placed E2F1/CDK1 transcriptional activation and PI3K/AKT signaling downstream of NLE1, defining an effector axis via rescue epistasis.

    Evidence NLE1 knockdown in NSCLC cells and xenografts with CDK1 overexpression and Akt activator rescue experiments

    PMID:36818671

    Open questions at the time
    • How NLE1 activates E2F1 transcription mechanistically not shown
    • Ordering of PI3K/AKT relative to E2F1/CDK1 not fully resolved
    • No direct NLE1-E2F1 interaction demonstrated
  5. 2024 Medium

    Direct interaction with Notch1 was demonstrated and exploited pharmacologically, establishing NLE1 as a druggable Notch-pathway component in brain tumor stem cells.

    Evidence Drug-target identification of edaravone binding NLE1, Notch reporter and self-renewal assays in BTSCs, and preclinical glioblastoma survival models

    PMID:39423824

    Open questions at the time
    • Precise biochemical NLE1-Notch1 binding interface not detailed
    • Whether edaravone-NLE1 binding is direct and specific not fully characterized
    • Relationship of the Notch1 complex to NLE1's protein-biosynthesis role unaddressed
  6. 2025 Low

    NLE1's pro-proliferative role was extended to hepatocellular carcinoma, reinforcing its broad pan-cancer growth function.

    Evidence CCK-8 and colony formation proliferation assays with CRISPR screen and multiomics enrichment analysis

    PMID:40625398

    Open questions at the time
    • Proliferation assays without pathway mechanistic follow-up
    • No direct partner or substrate identified in HCC
    • Correlative enrichment analysis only

Open questions

Synthesis pass · forward-looking unresolved questions
  • How NLE1's molecular role in protein biosynthesis mechanistically connects to its Notch1, PI3K/AKT, and E2F1/CDK1 functions remains unresolved.
  • No structural or biochemical definition of NLE1's molecular activity
  • No unifying model linking the biosynthetic and signaling functions
  • Direct substrates and stable complex composition unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Pathway
R-HSA-162582 Signal Transduction 3 R-HSA-1643685 Disease 2 R-HSA-392499 Metabolism of proteins 1
Partners
NOTCH1

Evidence

Reading pass · 8 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2022 NLE1 is upregulated in colorectal cancer upon SMAD4 loss via c-MYC occupying an E-box-containing region within the NLE1 promoter, thereby preventing TGFβ-mediated downregulation of NLE1. ChIP (c-MYC at NLE1 promoter E-box), patient-derived tumor organoids with ectopic c-MYC expression, TGFβ1 treatment Cancer research High 36219392
2022 NLE1 is limiting for de novo protein biosynthesis in colorectal cancer cells; its ablation activates p38/MAPK signaling, impairs autophagy (accumulation of p62- and LC3-positive structures), increases reactive oxygen species, and inhibits proliferation, migration, invasion, clonogenicity, and anchorage-independent growth. NLE1 knockout (CRISPR/KO) in colorectal cancer cell lines and organoids; protein synthesis assays; immunofluorescence for p62/LC3; ROS measurement; proliferation, migration, invasion, and colony formation assays Cancer research High 36219392
2022 NLE1 ablation impairs colorectal tumor growth in the colon and reduces primary tumor-derived liver metastasis in an endoscopy-guided orthotopic mouse transplantation model; deletion of TP53 further sensitizes NLE1-deficient cells to apoptosis. Orthotopic mouse transplantation model with NLE1 KO colorectal cancer cells; in vivo tumor growth and liver metastasis quantification; apoptosis assays with TP53 co-deletion Cancer research High 36219392
2024 NLE1 directly interacts with Notch1 to form a NLE1-Notch1 complex in brain tumor stem cells (BTSCs); the drug edaravone directly targets NLE1, impairs Notch signaling, alters stem cell marker expression, and sensitizes BTSCs to ionizing radiation-induced cell death. Global gene expression analysis, mechanistic drug-target identification of edaravone binding to NLE1, Notch pathway reporter assays, self-renewal/growth assays in BTSCs, preclinical glioblastoma mouse models with survival analysis Stem cell reports Medium 39423824
2017 NLE1 (notchless homolog 1) expression is reduced in the somatosensory cortex of WAG/Rij rats (genetic absence epilepsy model) compared to controls, coinciding with reduced Notch1 expression and altered neuron-to-glia ratio; acute Notch1 agonist (Jagged1) suppressed spike-and-wave discharges while Notch antagonist (DAPT) facilitated them, placing NLE1 as a modulator of the Notch1 signaling pathway in cortical development. Immunohistochemistry, qRT-PCR for NLE1 and Notch1 mRNA/protein in brain regions; in vivo EEG with pharmacological Notch1 agonist/antagonist administration Brain structure & function Medium 28210849
2023 NLE1 promotes NSCLC development by activating E2F1-mediated transcription of CDK1; PI3K/Akt signaling acts downstream of NLE1. CDK1 overexpression or Akt pathway activator treatment reversed NLE1 knockdown-induced NSCLC inhibition. NLE1 knockdown in NSCLC cell lines (in vitro loss-of-function); in vivo xenograft model; CDK1 rescue overexpression; Akt activator rescue experiment; cell growth, migration, and apoptosis assays Frontiers in oncology Medium 36818671
2021 NLE1 knockdown in melanoma cells inhibits proliferation, increases apoptotic sensitivity, and impairs migration in vitro, and inhibits tumor formation in vivo; the apoptosis induction requires apoptosis-related proteins and NLE1 activates the PI3K/AKT signaling pathway. shRNA knockdown of NLE1 in melanoma cell lines; proliferation, apoptosis, migration assays; mouse xenograft model; western blotting for PI3K/AKT pathway components and apoptosis-related proteins Experimental cell research Medium 34019907
2025 NLE1 (WD40 protein) promotes hepatocellular carcinoma cell proliferation as demonstrated by proliferation assays (CCK-8 and colony formation); enrichment analysis linked elevated NLE1 expression to proliferative gene programs. CCK-8 proliferation assay, colony formation assay, genome-wide CRISPR screen data, multiomics enrichment analysis Clinical Medicine Insights. Oncology Low 40625398

Source papers

Stage 0 corpus · 51 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2002 Bombesin receptor subtypes in human cancers: detection with the universal radioligand (125)I-[D-TYR(6), beta-ALA(11), PHE(13), NLE(14)] bombesin(6-14). Clinical cancer research : an official journal of the American Association for Cancer Research 281 11948125
2004 Effect of I.C.V. injection of AT4 receptor ligands, NLE1-angiotensin IV and LVV-hemorphin 7, on spatial learning in rats. Neuroscience 111 14980384
1987 Autoradiography of CCK receptors in the rat brain using [(3)H]Boc[Nle(28)(31)]CCK(27)-(33) and [(125)I]bolton-hunter CCK(8). Functional significance of subregional distributions. Neurochemistry international 72 20501122
2009 Low-density macroarray targeting non-locus of enterocyte effacement effectors (nle genes) and major virulence factors of Shiga toxin-producing Escherichia coli (STEC): a new approach for molecular risk assessment of STEC isolates. Applied and environmental microbiology 67 19880649
1997 Biological and conformational examination of stereochemical modifications using the template melanotropin peptide, Ac-Nle-c[Asp-His-Phe-Arg-Trp-Ala-Lys]-NH2, on human melanocortin receptors. Journal of medicinal chemistry 60 9171884
2004 FE200041 (D-Phe-D-Phe-D-Nle-D-Arg-NH2): A peripheral efficacious kappa opioid agonist with unprecedented selectivity. The Journal of pharmacology and experimental therapeutics 54 14993260
2014 64Cu- and 68Ga-labelled [Nle(14),Lys(40)(Ahx-NODAGA)NH2]-exendin-4 for pancreatic beta cell imaging in rats. Molecular imaging and biology 53 24101374
2012 Comprehensive immunohistochemical studies on canine necrotizing meningoencephalitis (NME), necrotizing leukoencephalitis (NLE), and granulomatous meningoencephalomyelitis (GME). Veterinary pathology 45 22262353
2003 Design of a new peptidomimetic agonist for the melanocortin receptors based on the solution structure of the peptide ligand, Ac-Nle-cyclo[Asp-Pro-DPhe-Arg-Trp-Lys]-NH(2). Bioorganic & medicinal chemistry letters 41 12824029
2000 VIP and the potent analog, stearyl-Nle(17)-VIP, induce proliferation of keratinocytes. FEBS letters 40 10858492
2012 Novel radiolabeled peptides for breast and prostate tumor PET imaging: (64)Cu/and (68)Ga/NOTA-PEG-[D-Tyr(6),βAla(11),Thi(13),Nle(14)]BBN(6-14). Bioconjugate chemistry 34 22770480
2022 SMAD4 Loss Induces c-MYC-Mediated NLE1 Upregulation to Support Protein Biosynthesis, Colorectal Cancer Growth, and Metastasis. Cancer research 33 36219392
1993 Anti-Ro(SS-A) HLA-DR3-positive women: the interrelationship between some ANA negative, SS, SCLE, and NLE mothers and SS/LE overlap female patients. The Journal of investigative dermatology 32 8423383
2010 The angiotensin IV analog Nle-Tyr-Leu-psi-(CH2-NH2)3-4-His-Pro-Phe (norleual) can act as a hepatocyte growth factor/c-Met inhibitor. The Journal of pharmacology and experimental therapeutics 31 20086056
1999 Parathyroid hormone (PTH)-(1-34), [Nle(8,18),Tyr34]PTH-(3-34) amide, PTH-(1-31) amide, and PTH-related peptide-(1-34) stimulate phosphatidylcholine hydrolysis in UMR-106 osteoblastic cells: comparison with effects of phorbol 12,13-dibutyrate. Endocrinology 30 9886817
2016 Pharmacodynamic evaluation of Lys5, MeLeu9, Nle10-NKA(4-10) prokinetic effects on bladder and colon activity in acute spinal cord transected and spinally intact rats. Naunyn-Schmiedeberg's archives of pharmacology 26 27889808
2005 Effect of MELANOTAN, [Nle(4), D-Phe(7)]-alpha-MSH, on melanin synthesis in humans with MC1R variant alleles. Peptides 25 16293341
1977 The gastrointestinal myoelectric response to 13-Nle-motilin infusion during interdigestive and digestive states in the conscious dog. Acta hepato-gastroenterologica 22 906776
2012 In vitro and in vivo pharmacological profile of PL-3994, a novel cyclic peptide (Hept-cyclo(Cys-His-Phe-d-Ala-Gly-Arg-d-Nle-Asp-Arg-Ile-Ser-Cys)-Tyr-[Arg mimetic]-NH(2)) natriuretic peptide receptor-A agonist that is resistant to neutral endopeptidase and acts as a bronchodilator. Pulmonary pharmacology & therapeutics 20 23154072
2018 Replacement of Arg with Nle and modified D-Phe in the core sequence of MSHs, Ac-His-D-Phe-Arg-Trp-NH2, leads to hMC1R selectivity and pigmentation. European journal of medicinal chemistry 19 29679901
2017 Developmental changes in Notch1 and NLE1 expression in a genetic model of absence epilepsy. Brain structure & function 17 28210849
2016 Low kidney uptake of GLP-1R-targeting, beta cell-specific PET tracer, 18F-labeled [Nle14,Lys40]exendin-4 analog, shows promise for clinical imaging. EJNMMI research 17 27957723
2012 A distinct regulatory sequence is essential for the expression of a subset of nle genes in attaching and effacing Escherichia coli. Journal of bacteriology 17 22904277
2017 A Macrocyclic Agouti-Related Protein/[Nle4,DPhe7]α-Melanocyte Stimulating Hormone Chimeric Scaffold Produces Subnanomolar Melanocortin Receptor Ligands. Journal of medicinal chemistry 14 28045525
1976 Potentiation between 13-Nle-motilin and acetylcholine on rabbit pyloric muscle in vitro. Scandinavian journal of gastroenterology. Supplement 14 1069362
2018 [Lys5,MeLeu9,Nle10]-NKA(4-10) Elicits NK2 Receptor-Mediated Micturition and Defecation, and NK1 Receptor-Mediated Emesis and Hypotension, in Conscious Dogs. The Journal of pharmacology and experimental therapeutics 13 29728445
1996 The use of topographical constraints in receptor mapping: investigation of the topographical requirements of the tryptophan 30 residue for receptor binding of Asp-Tyr-D-Phe-Gly-Trp-(N-Me)Nle-Asp-Phe-NH2 (SNF 9007), a cholecystokinin (26-33) analogue that binds to both CCK-B and delta-opioid receptors. Journal of medicinal chemistry 13 8831778
1984 Hydrolysis of the synthetic chromophoric hexapeptide Leu-Ser-Phe(NO2)-Nle-Ala-Leu-OMe catalyzed by bovine pepsin A. Dependence on pH and effect of enzyme phosphorylation level. Biochimica et biophysica acta 13 6437448
2017 Fluorescence- and Radiolabeling of [Lys4,Nle17,30]hPP Yields Molecular Tools for the NPY Y4 Receptor. Bioconjugate chemistry 12 28345900
2007 Discovery of Trp-Nle-Tyr-Met as a novel agonist for human formyl peptide receptor-like 1. Biochemical pharmacology 12 17517377
1976 Effects of in vivo administration of pentagastrin, secretin, and 13-Nle-Motilin on the in vitro incorporation of 14C-leucine into protein of human gastric mucosa. Scandinavian journal of gastroenterology 11 996430
1999 Importance of the aromatic residue at position 6 of [Nle(10)]neurokinin A(4-10) for binding to the NK-2 receptor and receptor activation. Journal of medicinal chemistry 10 10425111
1975 Protein synthesis in human gastric mucosa: effects of pentagastrin, secretin, and 13-nle-motilin. Acta hepato-gastroenterologica 10 1199672
2019 Chronic, Twice-Daily Dosing of an NK2 Receptor Agonist [Lys5,MeLeu9,Nle10]-NKA(4-10), Produces Consistent Drug-Induced Micturition and Defecation in Chronic Spinal Rats. Journal of neurotrauma 9 31642371
2003 Synthesis and binding characteristics of a novel enkephalin analogue, [3H]Tyr-D-Ala-Gly-Phe-D-Nle-Arg-Phe. Peptides 9 14706559
2023 Identification of NLE1/CDK1 axis as key regulator in the development and progression of non-small cell lung cancer. Frontiers in oncology 8 36818671
2021 Knockdown of NLE1 inhibits development of malignant melanoma in vitro and in vivo NLE1 promotes development of malignant melanoma. Experimental cell research 7 34019907
2020 [MeArg1, NLe10]-apelin-12: Optimization of solid-phase synthesis and evaluation of biological properties in vitro and in vivo. Peptides 7 32380198
2018 Colorectal and cardiovascular effects of [Lys5,MeLeu9,Nle10]-NKA(4-10) in anesthetized macaques. Naunyn-Schmiedeberg's archives of pharmacology 7 29858647
2016 Virulence traits and different nle profiles in cattle and human verotoxin-producing Escherichia coli O157:H7 strains from Argentina. Microbial pathogenesis 6 27914959
2024 Targeting glioblastoma with a brain-penetrant drug that impairs brain tumor stem cells via NLE1-Notch1 complex. Stem cell reports 5 39423824
2019 Prokinetic effects of the neurokinin NK2 receptor agonist [Lys5,MeLeu9,Nle10]-NKA(4-10) on bladder and colorectal activity in minipigs. Neuropeptides 5 31324387
1996 Effects of the novel cholecystokinin analogue Suc-Trp-N(Me)-Nle-Asp-Phe-NH2 on feeding and cortisol release in pigs. Pharmacology, biochemistry, and behavior 5 8728566
2018 Activation of bombesin receptor Subtype-3 by [D-Tyr6,β-Ala11,Phe13,Nle14]bombesin6-14 increased glucose uptake and lipogenesis in human and rat adipocytes. Molecular and cellular endocrinology 4 29402494
2022 [Lys5,MeLeu9,Nle10]-NKA(4-10) induces neurokinin 2 receptor mediated urination and defecation and neurokinin 1 receptor mediated flushing in rats: measured using the rapid detection voiding assay. Journal of basic and clinical physiology and pharmacology 3 36377965
2008 The micelle-associated 3D structures of Boc-Y(SO3)-Nle-G-W-Nle-D-2-phenylethylester (JMV-180) and CCK-8(s) share conformational elements of a calculated CCK1 receptor-bound model. Journal of medicinal chemistry 3 18540665
2022 Topological data analysis gives two folding paths in HP35(nle-nle), double mutant of villin headpiece subdomain. Scientific reports 2 35177744
2013 Effects of the Arg-Pro and Gly-Gly-Nle Moieties on Melanocortin-1 Receptor Binding Affinities of α-MSH Peptides. ACS medicinal chemistry letters 2 24349641
2025 WD40 Protein NLE1 as a Novel Diagnostic Biomarker Promoting Hepatocellular Carcinoma Proliferation. Clinical Medicine Insights. Oncology 1 40625398
2023 Colorectal and bladder prokinetic activity of [Lys5, MeLeu9, Nle10]-NKA(4-10) after intranasal or sublingual delivery in dogs. Canadian journal of physiology and pharmacology 1 36716436
2026 Preclinical assessment of 177Lu-DOTA-Pip-Nle-CycMSHhex: In vivo evaluation and human dosimetry estimation. Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine 0 42097084

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