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Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility

Cell migration requires a complex array of molecular events to promote protrusion at the front of motile cells. The scaffold protein LL5β interacts with the scaffold ERC1, and recruits it at plasma membrane–associated platforms that form at the front of migrating tumor cells. LL5 and ERC1 proteins s...

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Autores principales: Ribolla, Lucrezia Maria, Sala, Kristyna, Tonoli, Diletta, Ramella, Martina, Bracaglia, Lorenzo, Bonomo, Isabelle, Gonnelli, Leonardo, Lamarca, Andrea, Brindisi, Matteo, Pierattelli, Roberta, Provenzani, Alessandro, de Curtis, Ivan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10337942/
https://www.ncbi.nlm.nih.gov/pubmed/37437062
http://dx.doi.org/10.1371/journal.pone.0287670
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author Ribolla, Lucrezia Maria
Sala, Kristyna
Tonoli, Diletta
Ramella, Martina
Bracaglia, Lorenzo
Bonomo, Isabelle
Gonnelli, Leonardo
Lamarca, Andrea
Brindisi, Matteo
Pierattelli, Roberta
Provenzani, Alessandro
de Curtis, Ivan
author_facet Ribolla, Lucrezia Maria
Sala, Kristyna
Tonoli, Diletta
Ramella, Martina
Bracaglia, Lorenzo
Bonomo, Isabelle
Gonnelli, Leonardo
Lamarca, Andrea
Brindisi, Matteo
Pierattelli, Roberta
Provenzani, Alessandro
de Curtis, Ivan
author_sort Ribolla, Lucrezia Maria
collection PubMed
description Cell migration requires a complex array of molecular events to promote protrusion at the front of motile cells. The scaffold protein LL5β interacts with the scaffold ERC1, and recruits it at plasma membrane–associated platforms that form at the front of migrating tumor cells. LL5 and ERC1 proteins support protrusion during migration as shown by the finding that depletion of either endogenous protein impairs tumor cell motility and invasion. In this study we have tested the hypothesis that interfering with the interaction between LL5β and ERC1 may be used to interfere with the function of the endogenous proteins to inhibit tumor cell migration. For this, we identified ERC1(270–370) and LL5β(381–510) as minimal fragments required for the direct interaction between the two proteins. The biochemical characterization demonstrated that the specific regions of the two proteins, including predicted intrinsically disordered regions, are implicated in a reversible, high affinity direct heterotypic interaction. NMR spectroscopy further confirmed the disordered nature of the two fragments and also support the occurrence of interaction between them. We tested if the LL5β protein fragment interferes with the formation of the complex between the two full-length proteins. Coimmunoprecipitation experiments showed that LL5β(381–510) hampers the formation of the complex in cells. Moreover, expression of either fragment is able to specifically delocalize endogenous ERC1 from the edge of migrating MDA-MB-231 tumor cells. Coimmunoprecipitation experiments show that the ERC1-binding fragment of LL5β interacts with endogenous ERC1 and interferes with the binding of endogenous ERC1 to full length LL5β. Expression of LL5β(381–510) affects tumor cell motility with a reduction in the density of invadopodia and inhibits transwell invasion. These results provide a proof of principle that interfering with heterotypic intermolecular interactions between components of plasma membrane–associated platforms forming at the front of tumor cells may represent a new approach to inhibit cell invasion.
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spelling pubmed-103379422023-07-13 Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility Ribolla, Lucrezia Maria Sala, Kristyna Tonoli, Diletta Ramella, Martina Bracaglia, Lorenzo Bonomo, Isabelle Gonnelli, Leonardo Lamarca, Andrea Brindisi, Matteo Pierattelli, Roberta Provenzani, Alessandro de Curtis, Ivan PLoS One Research Article Cell migration requires a complex array of molecular events to promote protrusion at the front of motile cells. The scaffold protein LL5β interacts with the scaffold ERC1, and recruits it at plasma membrane–associated platforms that form at the front of migrating tumor cells. LL5 and ERC1 proteins support protrusion during migration as shown by the finding that depletion of either endogenous protein impairs tumor cell motility and invasion. In this study we have tested the hypothesis that interfering with the interaction between LL5β and ERC1 may be used to interfere with the function of the endogenous proteins to inhibit tumor cell migration. For this, we identified ERC1(270–370) and LL5β(381–510) as minimal fragments required for the direct interaction between the two proteins. The biochemical characterization demonstrated that the specific regions of the two proteins, including predicted intrinsically disordered regions, are implicated in a reversible, high affinity direct heterotypic interaction. NMR spectroscopy further confirmed the disordered nature of the two fragments and also support the occurrence of interaction between them. We tested if the LL5β protein fragment interferes with the formation of the complex between the two full-length proteins. Coimmunoprecipitation experiments showed that LL5β(381–510) hampers the formation of the complex in cells. Moreover, expression of either fragment is able to specifically delocalize endogenous ERC1 from the edge of migrating MDA-MB-231 tumor cells. Coimmunoprecipitation experiments show that the ERC1-binding fragment of LL5β interacts with endogenous ERC1 and interferes with the binding of endogenous ERC1 to full length LL5β. Expression of LL5β(381–510) affects tumor cell motility with a reduction in the density of invadopodia and inhibits transwell invasion. These results provide a proof of principle that interfering with heterotypic intermolecular interactions between components of plasma membrane–associated platforms forming at the front of tumor cells may represent a new approach to inhibit cell invasion. Public Library of Science 2023-07-12 /pmc/articles/PMC10337942/ /pubmed/37437062 http://dx.doi.org/10.1371/journal.pone.0287670 Text en © 2023 Ribolla et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Ribolla, Lucrezia Maria
Sala, Kristyna
Tonoli, Diletta
Ramella, Martina
Bracaglia, Lorenzo
Bonomo, Isabelle
Gonnelli, Leonardo
Lamarca, Andrea
Brindisi, Matteo
Pierattelli, Roberta
Provenzani, Alessandro
de Curtis, Ivan
Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility
title Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility
title_full Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility
title_fullStr Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility
title_full_unstemmed Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility
title_short Interfering with the ERC1–LL5β interaction disrupts plasma membrane–Associated platforms and affects tumor cell motility
title_sort interfering with the erc1–ll5β interaction disrupts plasma membrane–associated platforms and affects tumor cell motility
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10337942/
https://www.ncbi.nlm.nih.gov/pubmed/37437062
http://dx.doi.org/10.1371/journal.pone.0287670
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