{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Xlsx":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG.xlsx","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG.xlsx","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_91_meCpA.xlsx"],"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG_3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_91_meCpA_2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG_3.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG_2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_91_meCpA_1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG_2.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG_1.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG_1.raw"],"Mgf":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG_2.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG_3.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_91_meCpA_1.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG_3.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_61_meCpG_1.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG_1.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_91_meCpA_2.mgf","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD064826/Watson_Etal_2025_MeCP2_175bp_nucleosomes_position_1_meCpG_2.mgf"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["marcus.wilson@ed.ac.uk"],"submitter":["Christos Spanos"],"technology_type":["Mass Spectrometry","Chemical cross-linking coupled with mass spectrometry proteomics"],"software":["Not available"],"submitter_keywords":["Mecp2","Dna methylation","Chromatin","Cross-linking mass spectrometry","Nucleosomes"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD064826"],"tissue":["Cell Culture"],"sample_protocol":["Complexes of MeCP2 bound to methylated 175bp nucleosomes (2.5:1 molar ratio) were crosslinked using a concentration range of photo-reactive sulfo-SDA (sulfosuccinimidyl 4,4'-azipentanoate) (Thermo Fisher) in a w/w ratio of 1:0.25-1.5 (nucleosome: sulfo-SDA) in crosslink buffer [20 mM HEPES pH7.5, 250 mM NaCl, 1 mM EDTA, 1 mM DTT]. A total reaction volume of 30 µl, containing 7.5 µg of nucleosome, was incubated for 2 hrs on ice prior to UV irradiation at 365nm in a CL-1000L UV cross-linker (Spectrum) for 20 mins. The reaction was immediately quenched with 40 mM ammonium bicarbonate. The crosslinked sample was separated on an 3-8% gradient nuPAGE gel (Invitrogen) and bands running at a higher molecular weight than MeCP2 were excised. Protein gel bands were reduced with 10 mM TCEP for 30min at 37°C, alkylated with 55 mM iodoacetamide for 20min at room temperature and digested using 13 ng.μl-1 trypsin (Promega) overnight at 37°C. Digested peptides were desalted using C18-StageTips(Rappsilber et al., 2007) for LC-MS/MS analysis. LC-MS/MS analysis was performed using Orbitrap Fusion Lumos (Thermo Fisher Scientific) with a “high/high” acquisition strategy. The peptide separation was carried out on an EASY-Spray column (50 cm × 75 μm i.d., PepMap C18, 2 μm particles, 100 Å pore size, Thermo Fisher Scientific). Mobile phase A consisted of water and 0.1% v/v formic acid. Mobile phase B consisted of 80% v/v acetonitrile and 0.1% v/v formic acid. Peptides were loaded at a flow rate of 0.3 μl/min and eluted at 0.25 μl/min using a linear gradient going from 2% mobile phase B to 40% mobile phase B over 139 min (each sample has been running three time with different gradient), followed by a linear increase from 40% to 95% mobile phase B in 11 min (160 min total run time). The eluted peptides were directly introduced into the mass spectrometer. MS data were acquired in the data-dependent mode with 3 seconds acquisition cycle. Precursor spectrum was recorded in the Orbitrap with a resolution of 120,000. The ions with a precursor charge state between 3+ and 8+ were isolated with a window size of 1.6 m/z and fragmented using high-energy collision dissociation (HCD) with collision energy 30. The fragmentation spectra were recorded in the Orbitrap with a resolution of 30,000. Dynamic exclusion was enabled with single repeat count and 60 seconds exclusion duration."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Peak lists were generated with ProteoWizard ver. 3.0.24283 (Chambers et al, 2012).Cross-linked peptides were matched to spectra using Xi software (version 1.8.4); Mendes et al., 2018) with in-search assignment of monoisotopic peaks (Lenz et al., 2018). MS1 accuracy was set to 3 ppm and MS2 accuracy to 10 ppm. Trypsin was the protease of choice allowing four missed cleavages and SDA was chosen from the crosslinker list. Carbamidomethylation of cysteine was chosen as a fixed modification and oxidation of methionine was chosen as variable modification. TR False discovery rate was computed using XiFDR and results reported at 1% residue level false discovery rate (Fischer and Rappsilber, 2017). Our in-house protein database was used for the searches, containing MeCP2, and histones H2A, H2B, H3.1 and H4 (all Homo sapiens)."],"omics_type":["Proteomics"],"labhead":["Dr. Marcus Wilson"],"instrument_platform":[""],"labhead_affiliation":["University of Edinburgh"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["spanos.christos@gmail.com"],"publication":["41997957 Watson JA, Alexander-Howden BK, Hall TS, Wear MA, McGhie F, Clifford G, Wapenaar H, Zou J, Bird A, Wilson MD. MeCP2 requires interactions with nucleosome linker DNA to read chromatin DNA methylation. Nat Commun. 2026 17(1):5374 10.1038/s41467-026-71741-0"],"submitter_affiliation":["University of Edinburgh"],"submitter_country":["United Kingdom"],"pubmed_abstract":["Methyl-CpG-binding protein 2 (MeCP2) is a clinically important epigenetic reader that is essential for neuronal function, but how it binds methylated DNA within the protein-DNA complexes that comprise chromatin is unclear. Using designer nucleosomes, we observe that MeCP2 is able to engage methylated DNA at multiple sites on the nucleosome surface. Surprisingly, even methyl-cytosine placed in bent, histone-contacting, core nucleosomal DNA can be bound. However, we find that this ability requires interactions with inter-nucleosomal linker DNA. Nucleosome core DNA methylation reading involves regions of MeCP2 beyond its canonical methyl-CpG binding domain and we define a novel DNA-binding region in MeCP2 that is required for this function. We further demonstrate that histone H1 antagonises the MeCP2-nucleosome interactions by competing for linker DNA. Overall, our study reveals that MeCP2 gains access to methylated chromatinised DNA, independent of nucleosome structure, via essential nonspecific interactions with linker DNA."],"pubmed_title":["MeCP2 requires interactions with nucleosome linker DNA to read chromatin DNA methylation."],"pubmed_authors":["Watson James A JA, Alexander-Howden Beatrice K BK, Hall Theo S TS, Wear Martin A MA, McGhie Finlay F, Clifford Gillian G, Wapenaar Hannah H, Zou Juan J, Bird Adrian A, Wilson Marcus D MD"],"additional_accession":[]},"is_claimable":false,"name":"MeCP2 requires interactions with nucleosome linker DNA to read chromatin DNA methylation","description":"ethyl-CpG-binding protein 2 (MeCP2) is an epigenetic reader essential for neuronal function, but how it binds DNA methylation within chromatin is still unclear. Using designer nucleosomes we observe that MeCP2 preferentially binds DNA methylation positioned at multiple sites around the nucleosome. Surprisingly, MeCP2 can even bind methylated DNA in bent core nucleosomal DNA.  However, this activity requires additional DNA linker interactions, using regions outside of the canonical methyl binding domain. We mapped a novel DNA-binding region in MeCP2 required for this function. Furthermore, we find Histone H1 antagonises the MeCP2-nucleosome interaction through linker binding competition. Overall this reveals that MeCP2 combines nonspecific, but essential, interactions with linker DNA to aid specific binding to nucleosomal methylated DNA, independent of nucleosome structure. Our findings reveal in molecular detail how this clinically important protein interacts with chromatin and they identify novel domain that contributes to full MeCP2 function.","dates":{"publication":"2026-06-29","submission":"2025-06-10"},"accession":"PXD064826","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:108018","NEWT:377960","NEWT:1131","NEWT:35795","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NEWT:1334565","NEWT:625351","NEWT:258594","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:247480","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:2315688","NEWT:592321","NEWT:1073882","NEWT:6819","NEWT:46429","NEWT:475174","NEWT:111205","NEWT:1599","NEWT:428386","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:32264","NEWT:12215","NEWT:1590","NEWT:10034","NEWT:33114","NEWT:1596","NEWT:2736291","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:294671","NEWT:376619","NEWT:1748418","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:3767","NEWT:1589","NEWT:135622","NEWT:4615","NEWT:672127","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656061","NEWT:37965","NEWT:656064","NEWT:1582","NEWT:3760","NEWT:1584","NEWT:2086595","NEWT:36630","NEWT:1585","NEWT:383379","NEWT:381198","NEWT:418106","NCBITaxon:12721","NEWT:166340","NEWT:10029","NEWT:913645","NEWT:2492962","NEWT:641809","NEWT:44250","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:3357","NEWT:111225","NEWT:7719","NEWT:29073","NEWT:5530","NEWT:868565","NEWT:135674","NEWT:1562698","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:2767846","NEWT:108061","NEWT:60711","NEWT:2020713","NEWT:224308","NEWT:3347","NEWT:49928","NEWT:3349","NEWT:160621","NEWT:2502409","NEWT:212790","NEWT:110368","NEWT:55350","NEWT:1916073","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:468822","NEWT:1912919","NEWT:410658","NEWT:5516","NEWT:44688","NEWT:44689","NEWT:370038","NEWT:498211","NEWT:345076","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:1310173","NEWT:200795","NEWT:342808","NEWT:2579791","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:260499","NEWT:410661","NEWT:329854","NEWT:1140","NCBITaxon:2157","NEWT:2066563","NEWT:2649329","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:150808","NEWT:709964","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:156974","NEWT:3714","NCBITaxon:37162","NEWT:270643","NEWT:3715","NEWT:1538","NEWT:2065263","NEWT:199225","NEWT:177437","NEWT:717966","NEWT:9014","NEWT:10891","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:29022","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:176597","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:9006","NEWT:33548","NEWT:52299","NEWT:1883426","NEWT:8165","NEWT:96794","NEWT:10401","NEWT:10886","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:926571","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:1955249","NEWT:215451","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:2714896","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:1608994","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:329032","NEWT:56636","NEWT:2709072","NEWT:2499525","NEWT:55307","NEWT:1122981","NEWT:2853422","NEWT:1122984","NEWT:487","NEWT:1538553","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:45972","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:2836","NEWT:376686","NEWT:237610","NEWT:95486","EFO:0001352","NEWT:219809","NEWT:219808","NEWT:119562","NEWT:1883446","NEWT:31345","NEWT:1233435","NEWT:118223","NEWT:11757","NEWT:141469","NEWT:987059","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:1509","NEWT:491","NEWT:460523","NEWT:45954","NEWT:1578","NEWT:235443","NEWT:1579","NEWT:1757312","NEWT:85694","NEWT:109786","NEWT:1922344","NEWT:108458","NEWT:5936","NEWT:85692","NEWT:321967","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:279010","NEWT:291146","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:415978","NEWT:235455","NEWT:1255228","NEWT:410289","NEWT:191701","NEWT:373153","NEWT:1640278","NEWT:9054","NEWT:472","NEWT:940825","NEWT:29459","NEWT:529507","NEWT:1071661","NEWT:1380096","NEWT:28128","NEWT:470","NEWT:41122","NEWT:5911","NEWT:1328300","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:911022","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:717989","NEWT:33952","NEWT:1255232","NEWT:11723","NEWT:445","NEWT:153009","NEWT:1004879","NEWT:261756","NEWT:166314","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:29488","NEWT:1871049","NEWT:215402","NEWT:1547","NEWT:1290454","NEWT:3726","NEWT:906914","NEWT:81231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