Cargando…

A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis

KRAS is a powerful oncogene responsible for the development of many cancers. Despite the great progress in understanding its function during the last decade, the study of KRAS expression, subcellular localization, and post-translational modifications remains technically challenging. Accordingly, man...

Descripción completa

Detalles Bibliográficos
Autores principales: Assi, Mohamad, Pirlot, Boris, Stroobant, Vincent, Thissen, Jean-Paul, Jacquemin, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7504708/
https://www.ncbi.nlm.nih.gov/pubmed/32887255
http://dx.doi.org/10.3390/ijms21176361
_version_ 1783584685997162496
author Assi, Mohamad
Pirlot, Boris
Stroobant, Vincent
Thissen, Jean-Paul
Jacquemin, Patrick
author_facet Assi, Mohamad
Pirlot, Boris
Stroobant, Vincent
Thissen, Jean-Paul
Jacquemin, Patrick
author_sort Assi, Mohamad
collection PubMed
description KRAS is a powerful oncogene responsible for the development of many cancers. Despite the great progress in understanding its function during the last decade, the study of KRAS expression, subcellular localization, and post-translational modifications remains technically challenging. Accordingly, many facets of KRAS biology are still unknown. Antibodies could be an effective and easy-to-use tool for in vitro and in vivo research on KRAS. Here, we generated a novel rabbit polyclonal antibody that allows immunolabeling of cells and tissues overexpressing KRAS. Cell transfection experiments with expression vectors for the members of the RAS family revealed a preferential specificity of this antibody for KRAS. In addition, KRAS was sensitively detected in a mouse tissue electroporated with an expression vector. Interestingly, our antibody was able to detect endogenous forms of unprenylated (immature) and prenylated (mature) KRAS in mouse organs. We found that KRAS prenylation was increased ex vivo and in vivo in a model of KRAS(G12D)-driven tumorigenesis, which was concomitant with an induction of expression of essential KRAS prenylation enzymes. Therefore, our tool helped us to put the light on new regulations of KRAS activation during cancer initiation. The use of this tool by the RAS community could contribute to discovering novel aspects of KRAS biology.
format Online
Article
Text
id pubmed-7504708
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-75047082020-09-26 A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis Assi, Mohamad Pirlot, Boris Stroobant, Vincent Thissen, Jean-Paul Jacquemin, Patrick Int J Mol Sci Article KRAS is a powerful oncogene responsible for the development of many cancers. Despite the great progress in understanding its function during the last decade, the study of KRAS expression, subcellular localization, and post-translational modifications remains technically challenging. Accordingly, many facets of KRAS biology are still unknown. Antibodies could be an effective and easy-to-use tool for in vitro and in vivo research on KRAS. Here, we generated a novel rabbit polyclonal antibody that allows immunolabeling of cells and tissues overexpressing KRAS. Cell transfection experiments with expression vectors for the members of the RAS family revealed a preferential specificity of this antibody for KRAS. In addition, KRAS was sensitively detected in a mouse tissue electroporated with an expression vector. Interestingly, our antibody was able to detect endogenous forms of unprenylated (immature) and prenylated (mature) KRAS in mouse organs. We found that KRAS prenylation was increased ex vivo and in vivo in a model of KRAS(G12D)-driven tumorigenesis, which was concomitant with an induction of expression of essential KRAS prenylation enzymes. Therefore, our tool helped us to put the light on new regulations of KRAS activation during cancer initiation. The use of this tool by the RAS community could contribute to discovering novel aspects of KRAS biology. MDPI 2020-09-02 /pmc/articles/PMC7504708/ /pubmed/32887255 http://dx.doi.org/10.3390/ijms21176361 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Assi, Mohamad
Pirlot, Boris
Stroobant, Vincent
Thissen, Jean-Paul
Jacquemin, Patrick
A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis
title A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis
title_full A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis
title_fullStr A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis
title_full_unstemmed A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis
title_short A Novel KRAS Antibody Highlights a Regulation Mechanism of Post-Translational Modifications of KRAS during Tumorigenesis
title_sort novel kras antibody highlights a regulation mechanism of post-translational modifications of kras during tumorigenesis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7504708/
https://www.ncbi.nlm.nih.gov/pubmed/32887255
http://dx.doi.org/10.3390/ijms21176361
work_keys_str_mv AT assimohamad anovelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT pirlotboris anovelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT stroobantvincent anovelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT thissenjeanpaul anovelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT jacqueminpatrick anovelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT assimohamad novelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT pirlotboris novelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT stroobantvincent novelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT thissenjeanpaul novelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis
AT jacqueminpatrick novelkrasantibodyhighlightsaregulationmechanismofposttranslationalmodificationsofkrasduringtumorigenesis