Live visualization of a functional RET‐EGFP chimeric receptor in homozygous knock‐in mice

2021 ◽  
Author(s):  
Mukhamad Sunardi ◽  
Keisuke Ito ◽  
Hideki Enomoto
Keyword(s):  
Chimeric Receptor

Apoptosis Through CD95 (Fas/APO-1), but Not a CD40/CD95 Chimeric Receptor, Is Inhibited by Phorbol-12-Myristate-13-Acetate

1997 ◽  
Vol 16 (2) ◽  
pp. 197-205 ◽  
Author(s):  
FRITZ RUDERT ◽  
YUE WANG ◽  
ERICA LINDRIDGE ◽  
JAMES WATSON
Keyword(s):  
Chimeric Receptor

scholarly journals Efficient coupling with phosphatidylinositol 3-kinase, but not phospholipase C gamma or GTPase-activating protein, distinguishes ErbB-3 signaling from that of other ErbB/EGFR family members.

1994 ◽  
Vol 14 (1) ◽  
pp. 492-500 ◽  
Author(s):  
P Fedi ◽  
J H Pierce ◽  
P P di Fiore ◽  
M H Kraus
Keyword(s):  
Growth Factor ◽  
Phospholipase C ◽  
Recombinant Expression ◽  
Chimeric Receptor ◽  
Phosphatidylinositol 3 Kinase ◽  
Gtpase Activating Protein ◽  
Ligand Activation ◽  
Phospholipase C Gamma ◽  
Nih 3T3 ◽  
Phosphatidylinositol 3

Recombinant expression of a chimeric EGFR/ErbB-3 receptor in NIH 3T3 fibroblasts allowed us to investigate cytoplasmic events associated with ErbB-3 signal transduction upon ligand activation. An EGFR/ErbB-3 chimera was expressed on the surface of NIH 3T3 transfectants as two classes of receptors possessing epidermal growth factor (EGF) binding affinities comparable to those of the wild-type EGF receptor (EGFR). EGF induced autophosphorylation in vivo of the chimeric receptor and DNA synthesis of EGFR/ErbB-3 transfectants with a dose response similar to that of EGFR transfectants. However, the ErbB-3 and EGFR cytoplasmic domains exhibited striking differences in their interactions with several known tyrosine kinase substrates. We demonstrated strong association of phosphatidylinositol 3-kinase activity with the chimeric receptor upon ligand activation comparable in efficiency with that of the platelet-derived growth factor receptor, while the EGFR exhibited a 10- to 20-fold-lower efficiency in phosphatidylinositol 3-kinase recruitment. By contrast, both phospholipase C gamma and GTPase-activating protein failed to associate with or be phosphorylated by the ErbB-3 cytoplasmic domain under conditions in which they coupled with the EGFR. In addition, though certain signal transmitters, including Shc and GRB2, were recruited by both kinases, EGFR and ErbB-3 elicited tyrosine phosphorylation of distinct sets of intracellular substrates. Thus, our findings show that ligand activation of the ErbB-3 kinase triggers a cytoplasmic signaling pathway that hitherto is unique within this receptor subfamily.

In Vitro Comparison of Three Different Chimeric Receptor-modified Effector T-cell Populations for Leukemia Cell Therapy

2011 ◽  
Vol 34 (6) ◽  
pp. 469-479 ◽  
Author(s):  
Irene Pizzitola ◽  
Valentina Agostoni ◽  
Elisabetta Cribioli ◽  
Martin Pule ◽  
Raphael Rousseau ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Therapy ◽  
Leukemia Cell ◽  
Cell Populations ◽  
Chimeric Receptor ◽  
Effector T Cell ◽  
In Vitro Comparison

Efficient coupling with phosphatidylinositol 3-kinase, but not phospholipase C gamma or GTPase-activating protein, distinguishes ErbB-3 signaling from that of other ErbB/EGFR family members

1994 ◽  
Vol 14 (1) ◽  
pp. 492-500
Author(s):  
P Fedi ◽  
J H Pierce ◽  
P P di Fiore ◽  
M H Kraus
Keyword(s):  
Growth Factor ◽  
Phospholipase C ◽  
Recombinant Expression ◽  
Chimeric Receptor ◽  
Phosphatidylinositol 3 Kinase ◽  
Gtpase Activating Protein ◽  
Ligand Activation ◽  
Phospholipase C Gamma ◽  
Nih 3T3 ◽  
Phosphatidylinositol 3

Recombinant expression of a chimeric EGFR/ErbB-3 receptor in NIH 3T3 fibroblasts allowed us to investigate cytoplasmic events associated with ErbB-3 signal transduction upon ligand activation. An EGFR/ErbB-3 chimera was expressed on the surface of NIH 3T3 transfectants as two classes of receptors possessing epidermal growth factor (EGF) binding affinities comparable to those of the wild-type EGF receptor (EGFR). EGF induced autophosphorylation in vivo of the chimeric receptor and DNA synthesis of EGFR/ErbB-3 transfectants with a dose response similar to that of EGFR transfectants. However, the ErbB-3 and EGFR cytoplasmic domains exhibited striking differences in their interactions with several known tyrosine kinase substrates. We demonstrated strong association of phosphatidylinositol 3-kinase activity with the chimeric receptor upon ligand activation comparable in efficiency with that of the platelet-derived growth factor receptor, while the EGFR exhibited a 10- to 20-fold-lower efficiency in phosphatidylinositol 3-kinase recruitment. By contrast, both phospholipase C gamma and GTPase-activating protein failed to associate with or be phosphorylated by the ErbB-3 cytoplasmic domain under conditions in which they coupled with the EGFR. In addition, though certain signal transmitters, including Shc and GRB2, were recruited by both kinases, EGFR and ErbB-3 elicited tyrosine phosphorylation of distinct sets of intracellular substrates. Thus, our findings show that ligand activation of the ErbB-3 kinase triggers a cytoplasmic signaling pathway that hitherto is unique within this receptor subfamily.

Nanobody-based chimeric receptor gene integration in Jurkat cells mediated by PhiC31 integrase

2011 ◽  
Vol 317 (18) ◽  
pp. 2630-2641 ◽  
Author(s):  
Farnoush Jafari Iri-Sofla ◽  
Fatemeh Rahbarizadeh ◽  
Davoud Ahmadvand ◽  
Mohammad J. Rasaee
Keyword(s):  
Receptor Gene ◽  
Jurkat Cells ◽  
Chimeric Receptor ◽  
Phic31 Integrase ◽  
Gene Integration

scholarly journals An Inhibitory Region of the DNA-Binding Domain of Thyroid Hormone Receptor Blocks Hormone-Dependent Transactivation

1998 ◽  
Vol 12 (1) ◽  
pp. 34-44 ◽  
Author(s):  
Ying Liu ◽  
Akira Takeshita ◽  
Takashi Nagaya ◽  
Aria Baniahmad ◽  
William W. Chin ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Dna Binding ◽  
Ligand Binding ◽  
Transcriptional Activation ◽  
Thyroid Hormone Receptor ◽  
Binding Domain ◽  
Dna Binding Domain ◽  
Chimeric Receptor ◽  
Receptor System ◽  
Inhibitory Region

Abstract We have employed a chimeric receptor system in which we cotransfected yeast GAL4 DNA-binding domain/retinoid X receptor β ligand-binding domain chimeric receptor (GAL4RXR), thyroid hormone receptor-β (TRβ), and upstream activating sequence-reporter plasmids into CV-1 cells to study repression, derepression, and transcriptional activation. In the absence of T3, unliganded TR repressed transcription to 20% of basal level, and in the presence of T3, liganded TRβ derepressed transcription to basal level. Using this system and a battery of TRβ mutants, we found that TRβ/RXR heterodimer formation is necessary and sufficient for basal repression and derepression in this system. Additionally, an AF-2 domain mutant (E457A) mediated basal repression but not derepression, suggesting that interaction with a putative coactivator at this site may be critical for derepression. Interestingly, a mutant containing only the TRβ ligand binding domain (LBD) not only mediated derepression, but also stimulated transcriptional activation 10-fold higher than basal level. Studies using deletion and domain swap mutants localized an inhibitory region to the TRβ DNA-binding domain. Titration studies further suggested that allosteric changes promoting interaction with coactivators may account for enhanced transcriptional activity by LBD. In summary, our findings suggest that TR heterodimer formation with RXR is important for repression and derepression, and coactivator interaction with the AF-2 domain may be needed for derepression in this chimeric system. Additionally, there may be an inhibitory region in the DNA-binding domain, which reduces TR interaction with coactivators, and prevents full-length wild-type TRβ from achieving transcriptional activation above basal level in this chimeric receptor system.

scholarly journals Multiple regions within the cytoplasmic domains of the leukemia inhibitory factor receptor and gp130 cooperate in signal transduction in hepatic and neuronal cells.

1994 ◽  
Vol 14 (1) ◽  
pp. 138-146 ◽  
Author(s):  
H Baumann ◽  
A J Symes ◽  
M R Comeau ◽  
K K Morella ◽  
Y Wang ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Amino Acid ◽  
Leukemia Inhibitory Factor ◽  
Cytoplasmic Domain ◽  
Inhibitory Factor ◽  
Receptor Protein ◽  
Human Neuroblastoma Cell ◽  
Chimeric Receptor ◽  
Cytoplasmic Domains ◽  
Human Neuroblastoma

The receptor for leukemia inhibitory factor (LIFR), in combination with the signal-transducing subunit for interleukin-6-type cytokine receptors, gp130, and LIF, activates transcription of acute-phase plasma protein genes in human and rat hepatoma cells and the vasoactive intestinal peptide gene in a human neuroblastoma cell line. To identify the regions within the cytoplasmic domain of LIFR that initiate signal transduction independently of gp130, we constructed a chimeric receptor by linking the extracellular domain of the granulocyte colony-stimulating factor receptor (G-CSFR) to the transmembrane and cytoplasmic domain of human LIFR. The function of the chimeric receptor protein in transcriptional activation was assessed by G-CSF-mediated stimulation of cotransfected cytokine-responsive reporter gene constructs in hepatoma and neuroblastoma cells. By using the full-length cytoplasmic domain and mutants with progressive carboxy-terminal deletions, internal deletions, or point mutations, we identified the first 150 amino acid residues of LIFR as the minimal region necessary for signaling. The signaling reaction appears to involve a cooperativity between the first 70-amino-acid region containing the two sequence motifs conserved among hematopoietin receptors (box 1 and box 2) and a critical sequence between residues 141 and 150 (box 3). Analogous analyses of the cytoplasmic domains of G-CSFR and gp130 indicated similar arrangements of functional domains in these receptor subunits and the requirement of a box 3-related motif for signaling.

Abstract 1656: Vegf Induces vWF Release via A VEGFR2/Tyr1175-PLC- gamma1 Pathway

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Yan Xiong ◽  
Yingqing Huo ◽  
Chao Chen ◽  
Xiaofan Lu ◽  
Jincai Luo
Keyword(s):  
Growth Factor ◽  
Growth Factor Receptor ◽  
Initial Contact ◽  
Vegf Receptor ◽  
Chimeric Receptor ◽  
Vegf Receptors ◽  
Early Passage ◽  
Specific Storage ◽  
C Terminus

Many endothelial inflammatory and prothrombotic mediators are stored in and rapidly released from Weibel-Palade bodies (WPBs), endothelium-specific storage organelles upon stimulation. The von Willebrand factor (vWF), a major component inside WPBs, mediates the initial contact of platelets with the injured vessel wall and thus plays an important role in haemostasis and thrombosis. It has previously been shown that vascular endothelial growth factor (VEGF) triggers a rapid release of vWF. However, specific VEGF receptors and their potential downstream pathways involved in this process have not been carefully determined. To dissect the role of VEGF receptors in vWF release activation, we utilized two approaches: one is to use receptor-specific ligands and the other is to use a chimeric receptor approach. The ligands for VEGF receptor 2 (VEGFR2), but not VEGF receptor 1 (VEGFR1), stimulated vWF release. The predominant role of VEGFR2 in vWF release regulation was further confirmed by using a chimeric receptor approach in which the extracellular domain of the epidermal growth factor receptor was substituted for that of VEGFR1 or VEGFR2, and was then expressed in early-passage HUVECs. Further, the knockdown of phospholipase C-γ1 (PLCγ1) suppressed VEGF-triggered intracellular calcium increase and blocked VEGF-induced vWF release. In addition, the two products of PLCγ1 hydrolysis, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG), are both required for VEGF-induced vWF release. Finally, combined with point mutagenesis, the responsible binding sites for PLCγ1 and their importance in VEGFR2-activated vWF release have been determined. Point mutation of a single tyrosine residue Tyr1175, a putative binding site for PLCγ1 on the C-terminus, abolished VEGFR2-activated vWF release. This study presents the first evidence that the PLCγ1 is essential for VEGF-triggered vWF release mediated through a VEGFR2/Tyr1175 pathway.

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