Cargando…

Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1

BACKGROUND: Transformation by oncogene Ras overcomes TGF-β mediated growth inhibition in epithelial cells. However, it cooperates with each other to mediate epithelial to mesenchymal transition (EMT). The mechanism of how these two pathways interact with each other is controversial. METHODS: Molecul...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Sheng, Iaria, Josephine, Simpson, Richard J., Zhu, Hong-Jian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5850916/
https://www.ncbi.nlm.nih.gov/pubmed/29534718
http://dx.doi.org/10.1186/s12964-018-0223-4
_version_ 1783306307935141888
author Liu, Sheng
Iaria, Josephine
Simpson, Richard J.
Zhu, Hong-Jian
author_facet Liu, Sheng
Iaria, Josephine
Simpson, Richard J.
Zhu, Hong-Jian
author_sort Liu, Sheng
collection PubMed
description BACKGROUND: Transformation by oncogene Ras overcomes TGF-β mediated growth inhibition in epithelial cells. However, it cooperates with each other to mediate epithelial to mesenchymal transition (EMT). The mechanism of how these two pathways interact with each other is controversial. METHODS: Molecular techniques were used to engineer expression plasmids for Ras, SPRY, TGF-β receptors, type I and II and ubiquitin. Immunoprecipitation and western blots were employed to determine protein-protein interactions, preotein levels, protein phosphorylation while immunofluorecesent staining for molecular co-localization. TGF-β signalling activities is also determined by its luciferase reporter assay. Trans-well assays were used to measure cell migration and invasion. RESULTS: Ras interacts with the SPSB1’s SPRY domain to enhance TGF-β signaling. Ras interacts and colocalizes with the TGF-β type II receptor’s (TβRII) negative regulator SPSB1 on the cell membrane, consequently promoting SPSB1 protein degradation via enhanced mono- and di-ubiquitination. Reduced SPSB1 levels result in the stablization of TβRII, in turn the increase of receptor levels significantly enhance Smad2/3 phosphorylation and signaling. Importantly, forced expression of SPSB1 in Ras transformed cells suppresses TGF-β signaling and its mediated migration and invasion. CONCLUSION: Ras positively cooperates with TGF-β signaling by reducing the cellular protein levels of TβRII negative regualtor SPSB1. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12964-018-0223-4) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5850916
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-58509162018-03-21 Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1 Liu, Sheng Iaria, Josephine Simpson, Richard J. Zhu, Hong-Jian Cell Commun Signal Research BACKGROUND: Transformation by oncogene Ras overcomes TGF-β mediated growth inhibition in epithelial cells. However, it cooperates with each other to mediate epithelial to mesenchymal transition (EMT). The mechanism of how these two pathways interact with each other is controversial. METHODS: Molecular techniques were used to engineer expression plasmids for Ras, SPRY, TGF-β receptors, type I and II and ubiquitin. Immunoprecipitation and western blots were employed to determine protein-protein interactions, preotein levels, protein phosphorylation while immunofluorecesent staining for molecular co-localization. TGF-β signalling activities is also determined by its luciferase reporter assay. Trans-well assays were used to measure cell migration and invasion. RESULTS: Ras interacts with the SPSB1’s SPRY domain to enhance TGF-β signaling. Ras interacts and colocalizes with the TGF-β type II receptor’s (TβRII) negative regulator SPSB1 on the cell membrane, consequently promoting SPSB1 protein degradation via enhanced mono- and di-ubiquitination. Reduced SPSB1 levels result in the stablization of TβRII, in turn the increase of receptor levels significantly enhance Smad2/3 phosphorylation and signaling. Importantly, forced expression of SPSB1 in Ras transformed cells suppresses TGF-β signaling and its mediated migration and invasion. CONCLUSION: Ras positively cooperates with TGF-β signaling by reducing the cellular protein levels of TβRII negative regualtor SPSB1. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12964-018-0223-4) contains supplementary material, which is available to authorized users. BioMed Central 2018-03-13 /pmc/articles/PMC5850916/ /pubmed/29534718 http://dx.doi.org/10.1186/s12964-018-0223-4 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Liu, Sheng
Iaria, Josephine
Simpson, Richard J.
Zhu, Hong-Jian
Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1
title Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1
title_full Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1
title_fullStr Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1
title_full_unstemmed Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1
title_short Ras enhances TGF-β signaling by decreasing cellular protein levels of its type II receptor negative regulator SPSB1
title_sort ras enhances tgf-β signaling by decreasing cellular protein levels of its type ii receptor negative regulator spsb1
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5850916/
https://www.ncbi.nlm.nih.gov/pubmed/29534718
http://dx.doi.org/10.1186/s12964-018-0223-4
work_keys_str_mv AT liusheng rasenhancestgfbsignalingbydecreasingcellularproteinlevelsofitstypeiireceptornegativeregulatorspsb1
AT iariajosephine rasenhancestgfbsignalingbydecreasingcellularproteinlevelsofitstypeiireceptornegativeregulatorspsb1
AT simpsonrichardj rasenhancestgfbsignalingbydecreasingcellularproteinlevelsofitstypeiireceptornegativeregulatorspsb1
AT zhuhongjian rasenhancestgfbsignalingbydecreasingcellularproteinlevelsofitstypeiireceptornegativeregulatorspsb1