Cargando…

Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study

The causative gene of Fukuyama congenital muscular dystrophy (fukutin) is involved in formation of the basement membrane through glycosylation of alpha-dystroglycan. However, there are other proposed functions that have not been fully understood. Using cultured astrocytes (1321N1), we found nuclear...

Descripción completa

Detalles Bibliográficos
Autores principales: Okamura, Yukinori, Yamamoto, Tomoko, Tsukui, Ryota, Kato, Yoichiro, Shibata, Noriyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8622492/
https://www.ncbi.nlm.nih.gov/pubmed/34830034
http://dx.doi.org/10.3390/ijms222212153
_version_ 1784605706593239040
author Okamura, Yukinori
Yamamoto, Tomoko
Tsukui, Ryota
Kato, Yoichiro
Shibata, Noriyuki
author_facet Okamura, Yukinori
Yamamoto, Tomoko
Tsukui, Ryota
Kato, Yoichiro
Shibata, Noriyuki
author_sort Okamura, Yukinori
collection PubMed
description The causative gene of Fukuyama congenital muscular dystrophy (fukutin) is involved in formation of the basement membrane through glycosylation of alpha-dystroglycan. However, there are other proposed functions that have not been fully understood. Using cultured astrocytes (1321N1), we found nuclear localization of fukutin and a positive relationship between fukutin expression and cell proliferation. Among potential proteins regulating cell proliferation, we focused on cyclin D1, by reverse-transcription polymerase chain reaction, Western blotting, immunocytochemistry, enzyme-linked immunosorbent assay (ELISA), and sandwich ELISA. Expression of cyclin D1 was significantly downregulated by fukutin knockdown and significantly upregulated by fukutin overexpression. Moreover, fukutin was proven to bind to the activator protein-1 (AP-1) binding site of cyclin D1 promoter, as well as the AP-1 component c-Jun. The c-Jun phosphorylation status was not significantly influenced by knockdown or overexpression of fukutin. The present results provide in vitro evidence for a novel function of fukutin, which participates in cell proliferation by enhancing cyclin D1 expression through forming a complex with AP-1. It is likely that fukutin is a potential cofactor of AP-1.
format Online
Article
Text
id pubmed-8622492
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-86224922021-11-27 Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study Okamura, Yukinori Yamamoto, Tomoko Tsukui, Ryota Kato, Yoichiro Shibata, Noriyuki Int J Mol Sci Article The causative gene of Fukuyama congenital muscular dystrophy (fukutin) is involved in formation of the basement membrane through glycosylation of alpha-dystroglycan. However, there are other proposed functions that have not been fully understood. Using cultured astrocytes (1321N1), we found nuclear localization of fukutin and a positive relationship between fukutin expression and cell proliferation. Among potential proteins regulating cell proliferation, we focused on cyclin D1, by reverse-transcription polymerase chain reaction, Western blotting, immunocytochemistry, enzyme-linked immunosorbent assay (ELISA), and sandwich ELISA. Expression of cyclin D1 was significantly downregulated by fukutin knockdown and significantly upregulated by fukutin overexpression. Moreover, fukutin was proven to bind to the activator protein-1 (AP-1) binding site of cyclin D1 promoter, as well as the AP-1 component c-Jun. The c-Jun phosphorylation status was not significantly influenced by knockdown or overexpression of fukutin. The present results provide in vitro evidence for a novel function of fukutin, which participates in cell proliferation by enhancing cyclin D1 expression through forming a complex with AP-1. It is likely that fukutin is a potential cofactor of AP-1. MDPI 2021-11-10 /pmc/articles/PMC8622492/ /pubmed/34830034 http://dx.doi.org/10.3390/ijms222212153 Text en © 2021 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
Okamura, Yukinori
Yamamoto, Tomoko
Tsukui, Ryota
Kato, Yoichiro
Shibata, Noriyuki
Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study
title Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study
title_full Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study
title_fullStr Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study
title_full_unstemmed Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study
title_short Fukutin Protein Participates in Cell Proliferation by Enhancing Cyclin D1 Expression through Binding to the Transcription Factor Activator Protein-1: An In Vitro Study
title_sort fukutin protein participates in cell proliferation by enhancing cyclin d1 expression through binding to the transcription factor activator protein-1: an in vitro study
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8622492/
https://www.ncbi.nlm.nih.gov/pubmed/34830034
http://dx.doi.org/10.3390/ijms222212153
work_keys_str_mv AT okamurayukinori fukutinproteinparticipatesincellproliferationbyenhancingcyclind1expressionthroughbindingtothetranscriptionfactoractivatorprotein1aninvitrostudy
AT yamamototomoko fukutinproteinparticipatesincellproliferationbyenhancingcyclind1expressionthroughbindingtothetranscriptionfactoractivatorprotein1aninvitrostudy
AT tsukuiryota fukutinproteinparticipatesincellproliferationbyenhancingcyclind1expressionthroughbindingtothetranscriptionfactoractivatorprotein1aninvitrostudy
AT katoyoichiro fukutinproteinparticipatesincellproliferationbyenhancingcyclind1expressionthroughbindingtothetranscriptionfactoractivatorprotein1aninvitrostudy
AT shibatanoriyuki fukutinproteinparticipatesincellproliferationbyenhancingcyclind1expressionthroughbindingtothetranscriptionfactoractivatorprotein1aninvitrostudy