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Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket
The stem cell factor receptor (SCF) c-Kit plays a pivotal role in regulating cell proliferation and survival in many cell types. In particular, c-Kit is required for early amplification of erythroid progenitors, while it must disappear from cell surface for the cell entering the final steps of matur...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3634048/ https://www.ncbi.nlm.nih.gov/pubmed/23637779 http://dx.doi.org/10.1371/journal.pone.0060961 |
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author | D'allard, Diane Gay, Julie Descarpentries, Clotilde Frisan, Emilie Adam, Kevin Verdier, Frederique Floquet, Célia Dubreuil, Patrice Lacombe, Catherine Fontenay, Michaela Mayeux, Patrick Kosmider, Olivier |
author_facet | D'allard, Diane Gay, Julie Descarpentries, Clotilde Frisan, Emilie Adam, Kevin Verdier, Frederique Floquet, Célia Dubreuil, Patrice Lacombe, Catherine Fontenay, Michaela Mayeux, Patrick Kosmider, Olivier |
author_sort | D'allard, Diane |
collection | PubMed |
description | The stem cell factor receptor (SCF) c-Kit plays a pivotal role in regulating cell proliferation and survival in many cell types. In particular, c-Kit is required for early amplification of erythroid progenitors, while it must disappear from cell surface for the cell entering the final steps of maturation in an erythropoietin-dependent manner. We initially observed that imatinib (IM), an inhibitor targeting the tyrosine kinase activity of c-Kit concomitantly down-regulated the expression of c-Kit and accelerated the Epo-driven differentiation of erythroblasts in the absence of SCF. We investigated the mechanism by which IM or related masitinib (MA) induce c-Kit down-regulation in the human UT-7/Epo cell line. We found that the down-regulation of c-Kit in the presence of IM or MA was inhibited by a pre-incubation with methyl-β-cyclodextrin suggesting that c-Kit was internalized in the absence of ligand. By contrast to SCF, the internalization induced by TKI was independent of the E3 ubiquitin ligase c-Cbl. Furthermore, c-Kit was degraded through lysosomal, but not proteasomal pathway. In pulse-chase experiments, IM did not modulate c-Kit synthesis or maturation. Analysis of phosphotyrosine peptides in UT-7/Epo cells treated or not with IM show that IM did not modify overall tyrosine phosphorylation in these cells. Furthermore, we showed that a T670I mutation preventing the full access of IM to the ATP binding pocket, did not allow the internalization process in the presence of IM. Altogether these data show that TKI-induced internalization of c-Kit is linked to a modification of the integrity of ATP binding pocket. |
format | Online Article Text |
id | pubmed-3634048 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-36340482013-05-01 Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket D'allard, Diane Gay, Julie Descarpentries, Clotilde Frisan, Emilie Adam, Kevin Verdier, Frederique Floquet, Célia Dubreuil, Patrice Lacombe, Catherine Fontenay, Michaela Mayeux, Patrick Kosmider, Olivier PLoS One Research Article The stem cell factor receptor (SCF) c-Kit plays a pivotal role in regulating cell proliferation and survival in many cell types. In particular, c-Kit is required for early amplification of erythroid progenitors, while it must disappear from cell surface for the cell entering the final steps of maturation in an erythropoietin-dependent manner. We initially observed that imatinib (IM), an inhibitor targeting the tyrosine kinase activity of c-Kit concomitantly down-regulated the expression of c-Kit and accelerated the Epo-driven differentiation of erythroblasts in the absence of SCF. We investigated the mechanism by which IM or related masitinib (MA) induce c-Kit down-regulation in the human UT-7/Epo cell line. We found that the down-regulation of c-Kit in the presence of IM or MA was inhibited by a pre-incubation with methyl-β-cyclodextrin suggesting that c-Kit was internalized in the absence of ligand. By contrast to SCF, the internalization induced by TKI was independent of the E3 ubiquitin ligase c-Cbl. Furthermore, c-Kit was degraded through lysosomal, but not proteasomal pathway. In pulse-chase experiments, IM did not modulate c-Kit synthesis or maturation. Analysis of phosphotyrosine peptides in UT-7/Epo cells treated or not with IM show that IM did not modify overall tyrosine phosphorylation in these cells. Furthermore, we showed that a T670I mutation preventing the full access of IM to the ATP binding pocket, did not allow the internalization process in the presence of IM. Altogether these data show that TKI-induced internalization of c-Kit is linked to a modification of the integrity of ATP binding pocket. Public Library of Science 2013-04-23 /pmc/articles/PMC3634048/ /pubmed/23637779 http://dx.doi.org/10.1371/journal.pone.0060961 Text en © 2013 D'allard et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article D'allard, Diane Gay, Julie Descarpentries, Clotilde Frisan, Emilie Adam, Kevin Verdier, Frederique Floquet, Célia Dubreuil, Patrice Lacombe, Catherine Fontenay, Michaela Mayeux, Patrick Kosmider, Olivier Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket |
title | Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket |
title_full | Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket |
title_fullStr | Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket |
title_full_unstemmed | Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket |
title_short | Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket |
title_sort | tyrosine kinase inhibitors induce down-regulation of c-kit by targeting the atp pocket |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3634048/ https://www.ncbi.nlm.nih.gov/pubmed/23637779 http://dx.doi.org/10.1371/journal.pone.0060961 |
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