Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA

Abstract Whilst 5-methylcytosine (5mC) is a major epigenetic mark in the nuclear DNA in mammals, whether or not mitochondrial DNA (mtDNA) receives 5mC modification remains controversial. Herein, we exhaustively analysed mouse mtDNA using three methods that are based upon different principles for det...

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Autores principales:	Shigeru Matsuda, Takehiro Yasukawa, Yuriko Sakaguchi, Kenji Ichiyanagi, Motoko Unoki, Kazuhito Gotoh, Kei Fukuda, Hiroyuki Sasaki, Tsutomu Suzuki, Dongchon Kang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/292dcfe424644832b38fb2830dc1be9a
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spelling	oai:doaj.org-article:292dcfe424644832b38fb2830dc1be9a2021-12-02T15:08:41ZAccurate estimation of 5-methylcytosine in mammalian mitochondrial DNA10.1038/s41598-018-24251-z2045-2322https://doaj.org/article/292dcfe424644832b38fb2830dc1be9a2018-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-24251-zhttps://doaj.org/toc/2045-2322Abstract Whilst 5-methylcytosine (5mC) is a major epigenetic mark in the nuclear DNA in mammals, whether or not mitochondrial DNA (mtDNA) receives 5mC modification remains controversial. Herein, we exhaustively analysed mouse mtDNA using three methods that are based upon different principles for detecting 5mC. Next-generation bisulfite sequencing did not give any significant signatures of methylation in mtDNAs of liver, brain and embryonic stem cells (ESCs). Also, treatment with methylated cytosine-sensitive endonuclease McrBC resulted in no substantial decrease of mtDNA band intensities in Southern hybridisation. Furthermore, mass spectrometric nucleoside analyses of highly purified liver mtDNA preparations did not detect 5-methyldeoxycytidine at the levels found in the nuclear DNA but at a range of only 0.3–0.5% of deoxycytidine. Taken together, we propose that 5mC is not present at any specific region(s) of mtDNA and that levels of the methylated cytosine are fairly low, provided the modification occurs. It is thus unlikely that 5mC plays a universal role in mtDNA gene expression or mitochondrial metabolism.Shigeru MatsudaTakehiro YasukawaYuriko SakaguchiKenji IchiyanagiMotoko UnokiKazuhito GotohKei FukudaHiroyuki SasakiTsutomu SuzukiDongchon KangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-13 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shigeru Matsuda Takehiro Yasukawa Yuriko Sakaguchi Kenji Ichiyanagi Motoko Unoki Kazuhito Gotoh Kei Fukuda Hiroyuki Sasaki Tsutomu Suzuki Dongchon Kang Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA
description	Abstract Whilst 5-methylcytosine (5mC) is a major epigenetic mark in the nuclear DNA in mammals, whether or not mitochondrial DNA (mtDNA) receives 5mC modification remains controversial. Herein, we exhaustively analysed mouse mtDNA using three methods that are based upon different principles for detecting 5mC. Next-generation bisulfite sequencing did not give any significant signatures of methylation in mtDNAs of liver, brain and embryonic stem cells (ESCs). Also, treatment with methylated cytosine-sensitive endonuclease McrBC resulted in no substantial decrease of mtDNA band intensities in Southern hybridisation. Furthermore, mass spectrometric nucleoside analyses of highly purified liver mtDNA preparations did not detect 5-methyldeoxycytidine at the levels found in the nuclear DNA but at a range of only 0.3–0.5% of deoxycytidine. Taken together, we propose that 5mC is not present at any specific region(s) of mtDNA and that levels of the methylated cytosine are fairly low, provided the modification occurs. It is thus unlikely that 5mC plays a universal role in mtDNA gene expression or mitochondrial metabolism.
format	article
author	Shigeru Matsuda Takehiro Yasukawa Yuriko Sakaguchi Kenji Ichiyanagi Motoko Unoki Kazuhito Gotoh Kei Fukuda Hiroyuki Sasaki Tsutomu Suzuki Dongchon Kang
author_facet	Shigeru Matsuda Takehiro Yasukawa Yuriko Sakaguchi Kenji Ichiyanagi Motoko Unoki Kazuhito Gotoh Kei Fukuda Hiroyuki Sasaki Tsutomu Suzuki Dongchon Kang
author_sort	Shigeru Matsuda
title	Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA
title_short	Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA
title_full	Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA
title_fullStr	Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA
title_full_unstemmed	Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA
title_sort	accurate estimation of 5-methylcytosine in mammalian mitochondrial dna
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/292dcfe424644832b38fb2830dc1be9a
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Accurate estimation of 5-methylcytosine in mammalian mitochondrial DNA

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