No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos

The mechanisms responsible for active DNA demethylation remain elusive in Metazoa. A previous study that utilized zebrafish embryos provided a potent mechanism for active demethylation in which three proteins, AID, MBD4, and GADD45 are involved. We recently found age-dependent DNA hypomethylation in...

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Main Authors:	Shimoda, Nobuyoshi, Hirose, Kentaro, Kaneto, Reiya, Izawa, Toshiaki, Yokoi, Hayato, Hashimoto, Naohiro, Kikuchi, Yutaka
Format:	Online
Language:	English
Published:	Public Library of Science 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4275248/

id	pubmed-4275248
recordtype	oai_dc
spelling	pubmed-42752482014-12-31 No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos Shimoda, Nobuyoshi Hirose, Kentaro Kaneto, Reiya Izawa, Toshiaki Yokoi, Hayato Hashimoto, Naohiro Kikuchi, Yutaka Research Article The mechanisms responsible for active DNA demethylation remain elusive in Metazoa. A previous study that utilized zebrafish embryos provided a potent mechanism for active demethylation in which three proteins, AID, MBD4, and GADD45 are involved. We recently found age-dependent DNA hypomethylation in zebrafish, and it prompted us to examine if AID and MBD4 could be involved in the phenomenon. Unexpectedly, however, we found that most of the findings in the previous study were not reproducible. First, the injection of a methylated DNA fragment into zebrafish eggs did not affect either the methylation of genomic DNA, injected methylated DNA itself, or several loci tested or the expression level of aid, which has been shown to play a role in demethylation. Second, aberrant methylation was not observed at certain CpG islands following the injection of antisense morpholino oligonucleotides against aid and mbd4. Furthermore, we demonstrated that zebrafish MBD4 cDNA lacked a coding region for the methyl-CpG binding domain, which was assumed to be necessary for guidance to target regions. Taken together, we concluded that there is currently no evidence to support the proposed roles of AID and MBD4 in active demethylation in zebrafish embryos. Public Library of Science 2014-12-23 /pmc/articles/PMC4275248/ /pubmed/25536520 http://dx.doi.org/10.1371/journal.pone.0114816 Text en © 2014 Shimoda et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Shimoda, Nobuyoshi Hirose, Kentaro Kaneto, Reiya Izawa, Toshiaki Yokoi, Hayato Hashimoto, Naohiro Kikuchi, Yutaka
spellingShingle	Shimoda, Nobuyoshi Hirose, Kentaro Kaneto, Reiya Izawa, Toshiaki Yokoi, Hayato Hashimoto, Naohiro Kikuchi, Yutaka No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos
author_facet	Shimoda, Nobuyoshi Hirose, Kentaro Kaneto, Reiya Izawa, Toshiaki Yokoi, Hayato Hashimoto, Naohiro Kikuchi, Yutaka
author_sort	Shimoda, Nobuyoshi
title	No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos
title_short	No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos
title_full	No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos
title_fullStr	No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos
title_full_unstemmed	No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos
title_sort	no evidence for aid/mbd4-coupled dna demethylation in zebrafish embryos
description	The mechanisms responsible for active DNA demethylation remain elusive in Metazoa. A previous study that utilized zebrafish embryos provided a potent mechanism for active demethylation in which three proteins, AID, MBD4, and GADD45 are involved. We recently found age-dependent DNA hypomethylation in zebrafish, and it prompted us to examine if AID and MBD4 could be involved in the phenomenon. Unexpectedly, however, we found that most of the findings in the previous study were not reproducible. First, the injection of a methylated DNA fragment into zebrafish eggs did not affect either the methylation of genomic DNA, injected methylated DNA itself, or several loci tested or the expression level of aid, which has been shown to play a role in demethylation. Second, aberrant methylation was not observed at certain CpG islands following the injection of antisense morpholino oligonucleotides against aid and mbd4. Furthermore, we demonstrated that zebrafish MBD4 cDNA lacked a coding region for the methyl-CpG binding domain, which was assumed to be necessary for guidance to target regions. Taken together, we concluded that there is currently no evidence to support the proposed roles of AID and MBD4 in active demethylation in zebrafish embryos.
publisher	Public Library of Science
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4275248/
_version_	1613170254494564352

No Evidence for AID/MBD4-Coupled DNA Demethylation in Zebrafish Embryos

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