Fringe Glycosyltransferases Differentially Modulate Notch1 Proteolysis Induced by Delta1 and Jagged1

Fringe O-fucose-β1,3-N-acetylglucosaminyltransferases modulate Notch signaling by potentiating signaling induced by Delta-like ligands, while inhibiting signaling induced by Serrate/Jagged1 ligands. Based on binding studies, the differential effects of Drosophila fringe (DFng) on Notch signaling are thought to result from alterations in Notch glycosylation that enhance binding of Delta to Notch but reduce Serrate binding. Here, we report that expression of mammalian fringe proteins (Lunatic [LFng], Manic [MFng], or Radical [RFng] Fringe) increased Delta1 binding and activation of Notch1 signaling in 293T and NIH 3T3 cells. Although Jagged1-induced signaling was suppressed by LFng and MFng, RFng enhanced signaling induced by either Delta1 or Jagged1, underscoring the diversity of mammalian fringe glycosyltransferases in regulating signaling downstream of different ligand-receptor combinations. Interestingly, suppression of Jagged1-induced Notch1 signaling did not correlate with changes in Jagged1 binding as found for Delta1. Our data support the idea that fringe glycosylation increases Delta1 binding to potentiate signaling, but we propose that although fringe glycosylation does not reduce Jagged1 binding to Notch1, the resultant ligand–receptor interactions do not effectively promote Notch1 proteolysis required for activation of downstream signaling events.

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Augmentor α and β (FAM150) are ligands of the receptor tyrosine kinases ALK and LTK: Hierarchy and specificity of ligand–receptor interactions

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1520099112 ◽

2015 ◽

Vol 112 (52) ◽

pp. 15862-15867 ◽

Cited By ~ 64

Author(s):

Andrey V. Reshetnyak ◽

Phillip B. Murray ◽

Xiarong Shi ◽

Elizabeth S. Mo ◽

Jyotidarsini Mohanty ◽

...

Keyword(s):

Tyrosine Kinases ◽

Receptor Tyrosine Kinases ◽

Orphan Receptor ◽

3T3 Cells ◽

Cellular Functions ◽

Surface Receptors ◽

Anaplastic Lymphoma ◽

Disease States ◽

Receptor Interactions ◽

Nih 3T3

Receptor tyrosine kinases (RTKs) are a class of cell surface receptors that, upon ligand binding, stimulate a variety of critical cellular functions. The orphan receptor anaplastic lymphoma kinase (ALK) is one of very few RTKs that remain without a firmly established protein ligand. Here we present a novel cytokine, FAM150B, which we propose naming augmentor-α (AUG-α), as a ligand for ALK. AUG-α binds ALK with high affinity and activates ALK in cells with subnanomolar potency. Detailed binding experiments using cells expressing ALK or the related receptor leukocyte tyrosine kinase (LTK) demonstrate that AUG-α binds and robustly activates both ALK and LTK. We show that the previously established LTK ligand FAM150A (AUG-β) is specific for LTK and only weakly binds to ALK. Furthermore, expression of AUG-α stimulates transformation of NIH/3T3 cells expressing ALK, induces IL-3 independent growth of Ba/F3 cells expressing ALK, and is expressed in neuroblastoma, a cancer partly driven by ALK. These experiments reveal the hierarchy and specificity of two cytokines as ligands for ALK and LTK and set the stage for elucidating their roles in development and disease states.

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Binding of NCK to SOS and activation of ras-dependent gene expression.

Molecular and Cellular Biology ◽

10.1128/mcb.15.3.1169 ◽

1995 ◽

Vol 15 (3) ◽

pp. 1169-1174 ◽

Cited By ~ 67

Author(s):

Q Hu ◽

D Milfay ◽

L T Williams

Keyword(s):

Growth Factor ◽

Dominant Negative ◽

Tyrosine Kinase Receptors ◽

Nucleotide Exchange ◽

3T3 Cells ◽

Downstream Signaling ◽

Guanine Nucleotide Exchange ◽

Nucleotide Exchange Factor ◽

Nih 3T3 ◽

Sos Protein

NCK, an SH2- and SH3 domain-containing protein, becomes phosphorylated and associated with tyrosine kinase receptors upon growth factor stimulation. The sequence of NCK suggests that NCK functions as a linker between receptors and a downstream signaling molecule. To determine if NCK can mediate growth factor-stimulated responses, we measured the ability of NCK to activate the fos promoter. We found that in NIH 3T3 cells, NCK strongly activates this promoter. The effect of NCK on the fos promoter is enhanced by c-ras and blocked by dominant negative ras. We also found that NCK binds directly to the guanine nucleotide exchange factor SOS. This interaction is mediated by the SH3 domains of NCK. These findings suggest that NCK can regulate p21ras-dependent gene transcription through interaction with SOS protein.

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Differential Effects of Protein Kinase C Activation on Calcium Storage and Capacitative Calcium Entry in NIH 3T3 Cells

Journal of Biological Chemistry ◽

10.1074/jbc.271.35.21522 ◽

1996 ◽

Vol 271 (35) ◽

pp. 21522-21528 ◽

Cited By ~ 46

Author(s):

Carla M. Pedrosa Ribeiro ◽

James W. Putney

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Calcium Entry ◽

3T3 Cells ◽

Capacitative Calcium Entry ◽

Differential Effects ◽

Kinase C ◽

Calcium Storage ◽

Nih 3T3 ◽

Protein Kinase C Activation

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Lectin Binding Studies on RNA Tumor Virus-Infected Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100115250 ◽

1975 ◽

Vol 33 ◽

pp. 326-327

Author(s):

D. C. Hixson

Keyword(s):

Binding Sites ◽

Lectin Binding ◽

Culture Conditions ◽

3T3 Cells ◽

Binding Studies ◽

Con A ◽

Microscopic Studies ◽

Tumor Virus ◽

Infected Cells ◽

Cell Culture Conditions

The abilities of plant lectins to preferentially agglutinate malignant cells and to bind to specific monosaccharide or oligosaccharide sequences of glycoproteins and glycolipids make them a new and important biochemical probe for investigating alterations in plasma membrane structure which may result from malignant transformation. Electron and light microscopic studies have demonstrated clustered binding sites on surfaces of SV40-infected or tryp- sinized 3T3 cells when labeled with concanavalin A (con A). No clustering of con A binding sites was observed in normal 3T3 cells. It has been proposed that topological rearrangement of lectin binding sites into clusters enables con A to agglutinate SV40-infected or trypsinized 3T3 cells (1). However, observations by other investigators have not been consistent with this proposal (2) perhaps due to differences in reagents used, cell culture conditions, or labeling techniques. The present work was undertaken to study the lectin binding properties of normal and RNA tumor virus-infected cells and their associated viruses using lectins and ferritin-conjugated lectins of five different specificities.

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Induction of pseudofoci and inhibition of density-mediated neoplastic transformation by PMA in NIH 3T3 cells after short-term exposures

In Vitro Cellular & Developmental Biology - Animal ◽

10.1007/bf02639432 ◽

1995 ◽

Vol 31 (3) ◽

pp. 183-189 ◽

Cited By ~ 2

Author(s):

Andrew L. Rubin

Keyword(s):

Neoplastic Transformation ◽

3T3 Cells ◽

Short Term ◽

Nih 3T3 ◽

Nih 3T3 Cells

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Overexpression of protein kinase C-delta and -epsilon in NIH 3T3 cells induces opposite effects on growth, morphology, anchorage dependence, and tumorigenicity.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)53223-3 ◽

1993 ◽

Vol 268 (9) ◽

pp. 6090-6096 ◽

Cited By ~ 1

Author(s):

H. Mischak ◽

J.A. Goodnight ◽

W. Kolch ◽

G. Martiny-Baron ◽

C. Schaechtle ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

3T3 Cells ◽

Growth Morphology ◽

Protein Kinase C Delta ◽

Kinase C ◽

Nih 3T3 ◽

Nih 3T3 Cells

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CSIG-13. TRPM7 INDUCES TUMORIGENESIS AND STEMNESS THROUGH NOTCH ACTIVATION IN GLIOMA

Neuro-Oncology ◽

10.1093/neuonc/noaa215.125 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii30-ii30

Author(s):

Jingwei Wan ◽

Alyssa Guo ◽

Mingli Liu

Keyword(s):

Notch Signaling ◽

Glioma Cell ◽

Glioma Cells ◽

Inhibitory Effect ◽

Notch Signaling Pathway ◽

Normal Brain ◽

Diffuse Astrocytoma ◽

Notch1 Signaling ◽

Signaling Activation ◽

Proliferation And Invasion

Abstract Our group found that the inhibitory effect of TRPM7 on proliferation and invasion of human glioma cell is mediated by multiple mechanisms. TRPM7 regulates miR-28-5p expression, which suppresses cell proliferation and invasion in glioma cells by targeting Ras-related protein Rap1b. In particular, our group found that TRPM7 channels regulate glioma stem cell (GSC) growth/proliferation through STAT3 and Notch signaling. However, which Notch component(s) is crucial for its activity regulated by TRPM7, and its relationship with other GSC markers, such as CD133 and ALDH1, remain unclear. In the current project, we elucidate the mechanisms of TRMP7’s regulation of Notch signaling pathway that contribute to the development and progression of glioma and maintenance of self-renewal and tumorigenicity of GSC using multiple glioma cell lines (GC) with different molecular subtypes and GSCs derived from the GC lines. 1) We first analyzed TRPM7 expression using the Oncomine database (https://www.oncomine.org) and found that the TRPM7 mRNA expression is significantly increased in anaplastic astrocytoma, diffuse astrocytoma, and GBM patients compared to that in normal brain tissue controls. 2) TRPM7 is expressed in GBM, and its channel activity is correlated with Notch1 activation. Inhibition of TRPM7 downregulates Notch1 signaling, while upregulation of TRPM7 upregulates Notch1 signaling. 3) GSC markers, CD133 and ALDH1, are correlated with TRPM7 in GBM. 4) Targeting TRPM7 suppresses the growth and proliferation of glioma cells through G1/S arrests and apoptosis of glioma cells. 5) Targeting Notch1 suppresses the TRPM7-induced growth and proliferation of glioma cells, as well as the expression of GSC markers CD133 and ALDH1. In summary, TRPM7 is responsible for sustained Notch signaling activation, enhanced expression of GSC markers, and regulation of glioma stemness, which contribute to malignant glioma cell growth and invasion. Notch1 and ligand DII4 are key components that contribute GSC stemness.

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