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Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins

The determination of the protein’s intracellular localization is essential for understanding its biological function. Protein localization studies are mainly performed on primary and secondary vertebrate cell lines for which most protocols have been optimized. In spite of experimental difficulties,...

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Autores principales: Dominko, Kristina, Talajić, Antea, Radić, Martina, Vidaček, Nikolina Škrobot, Vlahoviček, Kristian, Bosnar, Maja Herak, Ćetković, Helena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9964533/
https://www.ncbi.nlm.nih.gov/pubmed/36827160
http://dx.doi.org/10.3390/md21020119
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author Dominko, Kristina
Talajić, Antea
Radić, Martina
Vidaček, Nikolina Škrobot
Vlahoviček, Kristian
Bosnar, Maja Herak
Ćetković, Helena
author_facet Dominko, Kristina
Talajić, Antea
Radić, Martina
Vidaček, Nikolina Škrobot
Vlahoviček, Kristian
Bosnar, Maja Herak
Ćetković, Helena
author_sort Dominko, Kristina
collection PubMed
description The determination of the protein’s intracellular localization is essential for understanding its biological function. Protein localization studies are mainly performed on primary and secondary vertebrate cell lines for which most protocols have been optimized. In spite of experimental difficulties, studies on invertebrate cells, including basal Metazoa, have greatly advanced. In recent years, the interest in studying human diseases from an evolutionary perspective has significantly increased. Sponges, placed at the base of the animal tree, are simple animals without true tissues and organs but with a complex genome containing many genes whose human homologs have been implicated in human diseases, including cancer. Therefore, sponges are an innovative model for elucidating the fundamental role of the proteins involved in cancer. In this study, we overexpressed human cancer-related proteins and their sponge homologs in human cancer cells, human fibroblasts, and sponge cells. We demonstrated that human and sponge MYC proteins localize in the nucleus, the RRAS2 in the plasma membrane, the membranes of the endolysosomal vesicles, and the DRG1 in the cell’s cytosol. Despite the very low transfection efficiency of sponge cells, we observed an identical localization of human proteins and their sponge homologs, indicating their similar cellular functions.
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spelling pubmed-99645332023-02-26 Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins Dominko, Kristina Talajić, Antea Radić, Martina Vidaček, Nikolina Škrobot Vlahoviček, Kristian Bosnar, Maja Herak Ćetković, Helena Mar Drugs Article The determination of the protein’s intracellular localization is essential for understanding its biological function. Protein localization studies are mainly performed on primary and secondary vertebrate cell lines for which most protocols have been optimized. In spite of experimental difficulties, studies on invertebrate cells, including basal Metazoa, have greatly advanced. In recent years, the interest in studying human diseases from an evolutionary perspective has significantly increased. Sponges, placed at the base of the animal tree, are simple animals without true tissues and organs but with a complex genome containing many genes whose human homologs have been implicated in human diseases, including cancer. Therefore, sponges are an innovative model for elucidating the fundamental role of the proteins involved in cancer. In this study, we overexpressed human cancer-related proteins and their sponge homologs in human cancer cells, human fibroblasts, and sponge cells. We demonstrated that human and sponge MYC proteins localize in the nucleus, the RRAS2 in the plasma membrane, the membranes of the endolysosomal vesicles, and the DRG1 in the cell’s cytosol. Despite the very low transfection efficiency of sponge cells, we observed an identical localization of human proteins and their sponge homologs, indicating their similar cellular functions. MDPI 2023-02-10 /pmc/articles/PMC9964533/ /pubmed/36827160 http://dx.doi.org/10.3390/md21020119 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dominko, Kristina
Talajić, Antea
Radić, Martina
Vidaček, Nikolina Škrobot
Vlahoviček, Kristian
Bosnar, Maja Herak
Ćetković, Helena
Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins
title Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins
title_full Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins
title_fullStr Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins
title_full_unstemmed Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins
title_short Transfection of Sponge Cells and Intracellular Localization of Cancer-Related MYC, RRAS2, and DRG1 Proteins
title_sort transfection of sponge cells and intracellular localization of cancer-related myc, rras2, and drg1 proteins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9964533/
https://www.ncbi.nlm.nih.gov/pubmed/36827160
http://dx.doi.org/10.3390/md21020119
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