{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T18:18:13Z","timestamp":1792001893313,"version":"4.4.0"},"reference-count":27,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T00:00:00Z","timestamp":1774051200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T00:00:00Z","timestamp":1774051200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100018246","name":"Lotte and Adolf Hotz Sprenger Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100018246","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2026,3,27]]},"abstract":"\n N6\u2010methyladenosine (m\n 6<\/jats:sup>\n A) is the most prevalent internal modification in eukaryotic messenger RNA. Dysregulation of m\n 6<\/jats:sup>\n A\u2010RNA signaling has been implicated in a wide range of human diseases. The N6\u2010methyladenosine modifications in DNA (6mA) is much rarer, and its role is still debated. Here, we report the first holo crystal structure of the m\n 6<\/jats:sup>\n A\u2010RNA reader YTHDC2. The 1.6\u2009\u00c5 resolution structure of YTHDC2 bound to the single\u2010strand (ss) hexanucleotide GG(6mA)CTA\u2010DNA shows an essentially identical binding mode of (6mA)CT\u2010DNA as (m\n 6<\/jats:sup>\n A)CU\u2010RNA in the other four human reader proteins YTHDC1 and YTHDF1\u20103. Comparative analysis of the binding of fluorescent\u2010labeled 6mA\u2010ssDNA and m\n 6<\/jats:sup>\n A\u2010RNA revealed that the five human m\n 6<\/jats:sup>\n A\u2010RNA readers exhibit slightly stronger binding affinity for 6mA\u2010modified DNA with a factor ranging from about 1.3 for YTHDC1 to 30 for YTHDC2. Given the similar affinity and the similar binding mode of 6mA\u2010ssDNA and m\n 6<\/jats:sup>\n A\u2010RNA, we set up to develop a fluorescence polarization (FP) binding assay that uses a fluorescent\u2010labeled 6mA\u2010containing ssDNA as probe. The DNA\u2010based FP assay shows high stability and robustness, making it suitable for high\u2010throughput screening applications. The assay provides a powerful and cost\u2010efficient platform to accelerate the discovery of small\u2010molecule modulators targeting m\n 6<\/jats:sup>\n A\u2010RNA reader proteins.\n <\/jats:p>","DOI":"10.1002\/cbic.202500897","type":"journal-article","created":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T18:05:43Z","timestamp":1774116343000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["A DNA\u2010Based Binding Assay for the m\n 6<\/sup>\n A\u2010RNA Reader Proteins"],"prefix":"10.1002","volume":"27","author":[{"given":"Rajiv Kumar","family":"Bedi","sequence":"first","affiliation":[{"name":"Department of Biochemistry University of Zurich Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2317-6792","authenticated-orcid":false,"given":"Amedeo","family":"Caflisch","sequence":"additional","affiliation":[{"name":"Department of Biochemistry University of Zurich Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,3,21]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature11112"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.05.003"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-019-0168-5"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.1432"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.687"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2012.10.015"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature12730"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2020105977"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1039\/D4CB00105B"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.8b05500"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aay3335"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201700424"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.10.006"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-020-0504-2"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2020.09.007"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/s44318-025-00512-2"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.0c01029"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsmedchemlett.2c00303"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.680389"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/nchembio.1654"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.05.012"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaa604"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsbiomedchemau.5c00099"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1177\/108705719900400206"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444911007773"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1107\/S2059798319011471"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444910007493"}],"container-title":["ChemBioChem"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202500897","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cbic.202500897","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cbic.202500897","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T10:16:01Z","timestamp":1775643361000},"score":1,"resource":{"primary":{"URL":"https:\/\/chemistry-europe.onlinelibrary.wiley.com\/doi\/10.1002\/cbic.202500897"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,21]]},"references-count":27,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2026,3,27]]}},"alternative-id":["10.1002\/cbic.202500897"],"URL":"https:\/\/doi.org\/10.1002\/cbic.202500897","archive":["Portico"],"relation":{},"ISSN":["1439-4227","1439-7633"],"issn-type":[{"value":"1439-4227","type":"print"},{"value":"1439-7633","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,21]]},"assertion":[{"value":"2025-11-17","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication 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