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Functional Comparison between Endogenous and Synthetic Notch Systems
[Image: see text] The Notch pathway converts receptor–ligand interactions at the cell surface into a transcriptional response in the receiver cell. In recent years, synthetic Notch systems (synNotch) that respond to different inputs and transduce different transcriptional responses have been enginee...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9594772/ https://www.ncbi.nlm.nih.gov/pubmed/36107643 http://dx.doi.org/10.1021/acssynbio.2c00247 |
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author | Khamaisi, Bassma Luca, Vincent C. Blacklow, Stephen C. Sprinzak, David |
author_facet | Khamaisi, Bassma Luca, Vincent C. Blacklow, Stephen C. Sprinzak, David |
author_sort | Khamaisi, Bassma |
collection | PubMed |
description | [Image: see text] The Notch pathway converts receptor–ligand interactions at the cell surface into a transcriptional response in the receiver cell. In recent years, synthetic Notch systems (synNotch) that respond to different inputs and transduce different transcriptional responses have been engineered. One class of synNotch systems uses antibody–antigen interactions at the cell surface to induce the proteolytic cleavage cascade of the endogenous Notch autoregulatory core and the consequent release of a synNotch intracellular domain (ICD), converting surface antigen detection into a cellular response. While the activation of endogenous Notch requires ubiquitylation and subsequent endocytosis of the ligand ICD, these synNotch systems do not seem to have such a requirement because the synNotch ligands completely lack an ICD. This observation raises questions about existing models for the synNotch activation mechanism. Here, we test how different structural and biochemical factors affect the dependence of endogenous and synthetic Notch activation on ligand ICD. We compare the behavior of antibody–antigen synNotch (aa-synNotch) to that of endogenous Notch, and to a synNotch system that uses rapamycin induced dimerization of FK506 binding protein (FKBP) and FKBP rapamycin binding (FRB) domaindimerization domains (ff-synNotch), which still requires a ligand ICD. We found that differences in receptor–ligand affinity, in the identity of the transmembrane domain, or in the presence or absence of extracellular epidermal growth factor repeats cannot explain the differences in ligand ICD requirement that distinguishes aa-synNotch from endogenous Notch or ff-synNotch. We also found that unlike endogenous Notch and ff-synNotch, the aa-synNotch system does not exhibit trans-endocytosis of the receptor extracellular domain into the sender cell. These findings suggest that the aa-synNotch systems bypass the ligand ICD requirement because antigen–antibody pairs are able to promote other adhesive cell–cell interactions that provide the mechanical tension needed for ligand activation. |
format | Online Article Text |
id | pubmed-9594772 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-95947722022-10-26 Functional Comparison between Endogenous and Synthetic Notch Systems Khamaisi, Bassma Luca, Vincent C. Blacklow, Stephen C. Sprinzak, David ACS Synth Biol [Image: see text] The Notch pathway converts receptor–ligand interactions at the cell surface into a transcriptional response in the receiver cell. In recent years, synthetic Notch systems (synNotch) that respond to different inputs and transduce different transcriptional responses have been engineered. One class of synNotch systems uses antibody–antigen interactions at the cell surface to induce the proteolytic cleavage cascade of the endogenous Notch autoregulatory core and the consequent release of a synNotch intracellular domain (ICD), converting surface antigen detection into a cellular response. While the activation of endogenous Notch requires ubiquitylation and subsequent endocytosis of the ligand ICD, these synNotch systems do not seem to have such a requirement because the synNotch ligands completely lack an ICD. This observation raises questions about existing models for the synNotch activation mechanism. Here, we test how different structural and biochemical factors affect the dependence of endogenous and synthetic Notch activation on ligand ICD. We compare the behavior of antibody–antigen synNotch (aa-synNotch) to that of endogenous Notch, and to a synNotch system that uses rapamycin induced dimerization of FK506 binding protein (FKBP) and FKBP rapamycin binding (FRB) domaindimerization domains (ff-synNotch), which still requires a ligand ICD. We found that differences in receptor–ligand affinity, in the identity of the transmembrane domain, or in the presence or absence of extracellular epidermal growth factor repeats cannot explain the differences in ligand ICD requirement that distinguishes aa-synNotch from endogenous Notch or ff-synNotch. We also found that unlike endogenous Notch and ff-synNotch, the aa-synNotch system does not exhibit trans-endocytosis of the receptor extracellular domain into the sender cell. These findings suggest that the aa-synNotch systems bypass the ligand ICD requirement because antigen–antibody pairs are able to promote other adhesive cell–cell interactions that provide the mechanical tension needed for ligand activation. American Chemical Society 2022-09-15 2022-10-21 /pmc/articles/PMC9594772/ /pubmed/36107643 http://dx.doi.org/10.1021/acssynbio.2c00247 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Khamaisi, Bassma Luca, Vincent C. Blacklow, Stephen C. Sprinzak, David Functional Comparison between Endogenous and Synthetic Notch Systems |
title | Functional Comparison between Endogenous and Synthetic
Notch Systems |
title_full | Functional Comparison between Endogenous and Synthetic
Notch Systems |
title_fullStr | Functional Comparison between Endogenous and Synthetic
Notch Systems |
title_full_unstemmed | Functional Comparison between Endogenous and Synthetic
Notch Systems |
title_short | Functional Comparison between Endogenous and Synthetic
Notch Systems |
title_sort | functional comparison between endogenous and synthetic
notch systems |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9594772/ https://www.ncbi.nlm.nih.gov/pubmed/36107643 http://dx.doi.org/10.1021/acssynbio.2c00247 |
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