{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T18:22:18Z","timestamp":1778523738211,"version":"3.51.4"},"reference-count":58,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2020,10,30]],"date-time":"2020-10-30T00:00:00Z","timestamp":1604016000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Czech Science Foundation","award":["18-07384S"],"award-info":[{"award-number":["18-07384S"]}]},{"name":"Strategie AV21, program Qualitas","award":["ICO: 68081707"],"award-info":[{"award-number":["ICO: 68081707"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>It has become evident that epitranscriptome events, mediated by specific enzymes, regulate gene expression and, subsequently, cell differentiation processes. We show that methyltransferase-like proteins METTL3\/METTL14 and N6-adenosine methylation (m6A) in RNAs are homogeneously distributed in embryonic hearts, and histone deacetylase (HDAC) inhibitors valproic acid and Trichostatin A (TSA) up-regulate METTL3\/METTL14 proteins. The levels of METTL3 in mouse adult hearts, isolated from male and female animals, were lower in the aorta and pulmonary trunks when compared with atria, but METT14 was up-regulated in the aorta and pulmonary trunk, in comparison with ventriculi. Aging caused METTL3 down-regulation in aorta and atria in male animals. Western blot analysis in differentiated mouse embryonic stem cells (mESCs), containing 10\u201330 percent of cardiomyocytes, showed METTL3\/METTL14 down-regulation, while the differentiation-induced increased level of METTL16 was observed in both wild type (wt) and HDAC1 depleted (dn) cells. In parallel, experimental differentiation in especially HDAC1 wild type cells was accompanied by depletion of m6A in RNA. Immunofluorescence analysis of individual cells revealed the highest density of METTL3\/METTL14 in \u03b1-actinin positive cardiomyocytes when compared with the other cells in the culture undergoing differentiation. In both wt and HDAC1 dn cells, the amount of METTL16 was also up-regulated in cardiomyocytes when compared to co-cultivated cells. Together, we showed that distinct anatomical regions of the mouse adult hearts are characterized by different levels of METTL3 and METTL14 proteins, which are changed during aging. Experimental cell differentiation was also accompanied by changes in METTL-like proteins and m6A in RNA; in particular, levels and distribution patterns of METTL3\/METTL14 proteins were different from the same parameters studied in the case of the METTL16 protein.<\/jats:p>","DOI":"10.3390\/ijms21218139","type":"journal-article","created":{"date-parts":[[2020,10,30]],"date-time":"2020-10-30T21:34:47Z","timestamp":1604093687000},"page":"8139","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["The Distinct Function and Localization of METTL3\/METTL14 and METTL16 Enzymes in Cardiomyocytes"],"prefix":"10.3390","volume":"21","author":[{"given":"Orazio Angelo","family":"Arcidiacono","sequence":"first","affiliation":[{"name":"Institute of Biophysics of the Czech Academy of Sciences, Kr\u00e1lovopolsk\u00e1 135, 61265 Brno, Czech Republic"},{"name":"Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 753\/5, 62500 Brno, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jana","family":"Krej\u010d\u00ed","sequence":"additional","affiliation":[{"name":"Institute of Biophysics of the Czech Academy of Sciences, Kr\u00e1lovopolsk\u00e1 135, 61265 Brno, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1299-0470","authenticated-orcid":false,"given":"Eva","family":"B\u00e1rtov\u00e1","sequence":"additional","affiliation":[{"name":"Institute of Biophysics of the Czech Academy of Sciences, Kr\u00e1lovopolsk\u00e1 135, 61265 Brno, Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"D259","DOI":"10.1093\/nar\/gkv1036","article-title":"RMBase: A resource for decoding the landscape of RNA modifications from high-throughput sequencing data","volume":"44","author":"Sun","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1038\/nrg3724","article-title":"Gene expression regulation mediated through reversible m 6 A RNA methylation","volume":"15","author":"Fu","year":"2014","journal-title":"Nat. Rev. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1101\/gad.262766.115","article-title":"RNA N6-methyladenosine methylation in post-transcriptional gene expression regulation","volume":"29","author":"Yue","year":"2015","journal-title":"Genes Dev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1038\/nature19342","article-title":"M6 A RNA methylation promotes XIST-mediated transcriptional repression","volume":"537","author":"Patil","year":"2016","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1038\/cr.2014.3","article-title":"Mammalian WTAP is a regulatory subunit of the RNA N6-methyladenosine methyltransferase","volume":"24","author":"Ping","year":"2014","journal-title":"Cell Res."},{"key":"ref_6","first-page":"4456","article-title":"N6-methyladenosine residues in an intron-specific region of prolactin pre-mRNA","volume":"10","author":"Carroll","year":"1990","journal-title":"Mol. Cell. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/0092-8674(76)90022-2","article-title":"Methyl labeling of HeLa cell hnRNA: A comparison with mRNA","volume":"7","author":"Jelinek","year":"1976","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1261\/rna.064063.117","article-title":"Interactions, localization, and phosphorylation of the m6A generating METTL3\u2013METTL14\u2013WTAP complex","volume":"24","author":"Weichmann","year":"2018","journal-title":"RNA"},{"key":"ref_9","first-page":"1233","article-title":"Purification and cDNA cloning of the AdoMet-binding subunit of the human mRNA (N6-adenosine)-methyltransferase","volume":"3","author":"Bokar","year":"1997","journal-title":"RNA"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1038\/nchembio.1432","article-title":"A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation","volume":"10","author":"Liu","year":"2014","journal-title":"Nat. Chem. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"109694","DOI":"10.1016\/j.biopha.2019.109694","article-title":"New sights in cancer: Component and function of N6-methyladenosine modification","volume":"122","author":"Hu","year":"2020","journal-title":"Biomed. Pharmacother."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/j.celrep.2014.05.048","article-title":"Perturbation of m6A writers reveals two distinct classes of mRNA methylation at internal and 5\u2032 sites","volume":"8","author":"Schwartz","year":"2014","journal-title":"Cell Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1101\/gad.309146.117","article-title":"Zc3h13\/Flacc is required for adenosine methylation by bridging the mRNA-binding factor RbM15\/spenito to the m6 a machinery component Wtap\/Fl(2)d","volume":"32","author":"Knuckles","year":"2018","journal-title":"Genes Dev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2004","DOI":"10.15252\/embr.201744940","article-title":"Human METTL16 is a N6 -methyladenosine (m 6 A) methyltransferase that targets pre-mRNAs and various non-coding RNAs","volume":"18","author":"Warda","year":"2017","journal-title":"EMBO Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1016\/j.cell.2017.05.003","article-title":"The U6 snRNA m6A methyltransferase METTL16 regulates SAM synthetase intron retention","volume":"169","author":"Pendleton","year":"2017","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1038\/nchembio.687","article-title":"N6-Methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO","volume":"7","author":"Jia","year":"2011","journal-title":"Nat. Chem. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.molcel.2012.10.015","article-title":"ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility","volume":"49","author":"Zheng","year":"2013","journal-title":"Mol. Cell"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Kurowski, M.A., Bhagwat, A.S., Papaj, G., and Bujnicki, J.M. (2003). Phylogenomic identification of five new human homologs of the DNA repair enzyme AlkB. BMC Genomics, 4.","DOI":"10.1186\/1471-2164-4-48"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"8885","DOI":"10.1002\/anie.201001242","article-title":"The AlkB domain of mammalian ABH8 catalyzes hydroxylation of 5-methoxycarbonylmethyluridine at the wobble position of tRNA","volume":"49","author":"Fu","year":"2010","journal-title":"Angew. Chemie Int. Ed."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1126\/science.1151710","article-title":"The obesity-associated FTO gene encodes a 2-oxoglutarate-dependent nucleic acid demethylase","volume":"318","author":"Gerken","year":"2007","journal-title":"Science"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1798","DOI":"10.1038\/ncomms2822","article-title":"FTO-mediated formation of N6-hydroxymethyladenosine and N 6-formyladenosine in mammalian RNA","volume":"4","author":"Fu","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.1126\/science.1210597","article-title":"Tet proteins can convert 5-methylcytosine to 5-formylcytosine and 5-carboxylcytosine","volume":"333","author":"Ito","year":"2011","journal-title":"Science"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1038\/nature09303","article-title":"Role of tet proteins in 5mC to 5hmC conversion, ES-cell self-renewal and inner cell mass specification","volume":"466","author":"Ito","year":"2010","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1126\/science.1170116","article-title":"Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1","volume":"324","author":"Tahiliani","year":"2009","journal-title":"Science"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1016\/j.cell.2013.10.026","article-title":"XRNA-methylation-dependent RNA processing controls the speed of the circadian clock","volume":"155","author":"Fustin","year":"2013","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1038\/nature14234","article-title":"N6 -methyladenosine-dependent RNA structural switches regulate RNA-protein interactions","volume":"518","author":"Liu","year":"2015","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1016\/j.molcel.2016.01.012","article-title":"Nuclear m6A reader YTHDC1 regulates mRNA splicing","volume":"61","author":"Xiao","year":"2016","journal-title":"Mol. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1038\/nature08697","article-title":"HnRNP proteins controlled by c-Myc deregulate pyruvate kinase mRNA splicing in cancer","volume":"463","author":"David","year":"2010","journal-title":"Nature"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1038\/nsmb.1838","article-title":"ICLIP reveals the function of hnRNP particles in splicing at individual nucleotide resolution","volume":"17","author":"Zarnack","year":"2010","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1038\/ncb2902","article-title":"N6 -methyladenosine modification destabilizes developmental regulators in embryonic stem cells","volume":"16","author":"Wang","year":"2014","journal-title":"Nat. Cell Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1038\/nature12730","article-title":"N6-methyladenosine-dependent regulation of messenger RNA stability","volume":"505","author":"Wang","year":"2014","journal-title":"Nature"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Svobodov\u00e1 Kova\u0159\u00edkov\u00e1, A., Stixov\u00e1, L., Kova\u0159\u00edk, A., Kom\u016frkov\u00e1, D., Legartov\u00e1, S., Fagherazzi, P., and B\u00e1rtov\u00e1, E. (2020). N6-Adenosine Methylation in RNA and a reduced m3G\/TMG level in non-coding RNAs appear at microirradiation-induced DNA lesions. Cells, 9.","DOI":"10.3390\/cells9020360"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1002","DOI":"10.1126\/science.1261417","article-title":"m6A mRNA methylation facilitates resolution of na\u00efve pluripotency toward differentiation","volume":"347","author":"Geula","year":"2015","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/j.stem.2014.09.019","article-title":"M6A RNA modification controls cell fate transition in mammalian embryonic stem cells","volume":"15","author":"Batista","year":"2014","journal-title":"Cell Stem Cell"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1016\/j.stem.2015.09.005","article-title":"Coordination of m6A mRNA methylation and gene transcription by ZFP217 regulates pluripotency and reprogramming","volume":"17","author":"Aguilo","year":"2015","journal-title":"Cell Stem Cell"},{"key":"ref_36","first-page":"E2047","article-title":"Hypoxia induces the breast cancer stem cell phenotype by HIF-dependent and ALKBH5-mediated m6A-demethylation of NANOG mRNA","volume":"113","author":"Zhang","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7913","DOI":"10.1128\/MCB.01220-06","article-title":"Negative and positive regulation of gene expression by mouse histone deacetylase 1","volume":"26","author":"Zupkovitz","year":"2006","journal-title":"Mol. Cell. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2672","DOI":"10.1093\/emboj\/21.11.2672","article-title":"Essential function of histone deacetylase 1 in proliferation control and CDK inhibitor repression","volume":"21","author":"Lagger","year":"2002","journal-title":"EMBO J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1790","DOI":"10.1101\/gad.1563807","article-title":"Histone deacetylases 1 and 2 redundantly regulate cardiac morphogenesis, growth, and contractility","volume":"21","author":"Montgomery","year":"2007","journal-title":"Genes Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/0925-4773(93)90015-P","article-title":"Embryonic stem cells differentiate in vitro into cardiomyocytes representing sinusnodal, atrial and ventricular cell types","volume":"44","author":"Maltsev","year":"1993","journal-title":"Mech. Dev."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Arcidiacono, O.A., Krej\u010d\u00ed, J., Such\u00e1nkov\u00e1, J., and B\u00e1rtov\u00e1, E. (2018). Deacetylation of histone H4 accompanying cardiomyogenesis is weakened in HDAC1-depleted ES cells. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19082425"},{"key":"ref_42","first-page":"532","article-title":"Troponin I serum concentration: A new marker of left ventricular hypertrophy in patients with essential hypertension","volume":"1","author":"Siciliano","year":"2000","journal-title":"Ital. Hear. J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1111\/j.1432-0436.1991.tb00255.x","article-title":"Pluripotent mouse embryonic stem cells are able to differentiate into cardiomyocytes expressing chronotropic responses to adrenergic and cholinergic agents and Ca2+ channel blockers","volume":"48","author":"Wobus","year":"1991","journal-title":"Differentiation"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1038\/s41593-017-0057-1","article-title":"N6-methyladenosine RNA modification regulates embryonic neural stem cell self-renewal through histone modifications","volume":"21","author":"Wang","year":"2018","journal-title":"Nat. Neurosci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1038\/s41586-019-1016-7","article-title":"Histone H3 trimethylation at lysine 36 guides m6A RNA modification co-transcriptionally","volume":"567","author":"Huang","year":"2019","journal-title":"Nature"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1016\/j.molcel.2018.08.004","article-title":"Methylation of structured RNA by the m6A writer METTL16 is essential for mouse embryonic development","volume":"71","author":"Mendel","year":"2018","journal-title":"Mol Cell."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Nance, D.J., Satterwhite, E.R., Bhaskar, B., Misra, S., Carraway, K.R., and Mansfield, K.D. (2020). Characterization of METTL16 as a cytoplasmic RNA binding protein. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0227647"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1016\/j.molcel.2018.07.025","article-title":"Structural basis for regulation of METTL16, an S-Adenosylmethionine Homeostasis Factor","volume":"71","author":"Doxtader","year":"2018","journal-title":"Mol. Cell"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1161\/CIRCULATIONAHA.118.036146","article-title":"The N-Methyladenosine mRNA Methylase METTL3 controls cardiac homeostasis and hypertrophy","volume":"139","author":"Dorn","year":"2019","journal-title":"Circulation"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Kmietczyk, V., Riechert, E., Kalinski, L., Boileau, E., Malovrh, E., Malone, B., Gorska, A., Hofmann, C., Varma, E., and J\u00fcrgensen, L. (2019). M 6 A-mRNA methylation regulates cardiac gene expression and cellular growth. Life Sci. Alliance, 2.","DOI":"10.26508\/lsa.201800233"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1002\/ejhf.1672","article-title":"Changes in m6A RNA methylation contribute to heart failure progression by modulating translation","volume":"22","author":"Berulava","year":"2020","journal-title":"Eur. J. Heart Fail."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1161\/CIRCULATIONAHA.118.033794","article-title":"FTO-dependent N-Methyladenosine regulates cardiac function during remodeling and repair","volume":"139","author":"Mathiyalagan","year":"2019","journal-title":"Circulation"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"6838","DOI":"10.1074\/jbc.M211762200","article-title":"The transcriptional co-activators CREB-binding protein (CBP) and p300 play a critical role in cardiac hypertrophy that is dependent on their histone acetyltransferase activity","volume":"278","author":"Gusterson","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"28930","DOI":"10.1074\/jbc.M303113200","article-title":"Dose-dependent blockade to cardiomyocyte hypertrophy by histone deacetylase inhibitors","volume":"278","author":"Antos","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"4123","DOI":"10.1073\/pnas.1015081108","article-title":"Histone deacetylase (HDAC) inhibitors attenuate cardiac hypertrophy by suppressing autophagy","volume":"108","author":"Cao","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1016\/j.yjmcc.2008.08.015","article-title":"Histone-deacetylase inhibition reverses atrial arrhythmia inducibility and fibrosis in cardiac hypertrophy independent of angiotensin","volume":"45","author":"Liu","year":"2008","journal-title":"J. Mol. Cell. Cardiol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1146\/annurev-pharmtox-010611-134712","article-title":"Therapeutic potential for HDAC inhibitors in the heart","volume":"52","author":"McKinsey","year":"2012","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"key":"ref_58","unstructured":"Mendis, S., and Puska, B.N. (2020, June 26). WHO | Global Atlas on Cardiovascular Disease Prevention and Control. Available online: http:\/\/www.who.int\/cardiovascular_diseases\/publications\/atlas_cvd\/en\/."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/21\/8139\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:27:22Z","timestamp":1760178442000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/21\/8139"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,30]]},"references-count":58,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2020,11]]}},"alternative-id":["ijms21218139"],"URL":"https:\/\/doi.org\/10.3390\/ijms21218139","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,30]]}}}