{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,17]],"date-time":"2025-10-17T14:13:13Z","timestamp":1760710393490,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2020,11,10]],"date-time":"2020-11-10T00:00:00Z","timestamp":1604966400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministerio de Econom\u00eda y Competitividad (Spain)","award":["BFU2016-78232-P"],"award-info":[{"award-number":["BFU2016-78232-P"]}]},{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["CPII13\/00017","PI15\/00663","PI18\/00349"],"award-info":[{"award-number":["CPII13\/00017","PI15\/00663","PI18\/00349"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010067","name":"Gobierno de Arag\u00f3n","doi-asserted-by":"publisher","award":["E45_17R","B25_17R"],"award-info":[{"award-number":["E45_17R","B25_17R"]}],"id":[{"id":"10.13039\/501100010067","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red en Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd)","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Methyl-CpG binding protein 2 (MeCP2) is a transcriptional regulator and a chromatin-binding protein involved in neuronal development and maturation. Loss-of-function mutations in MeCP2 result in Rett syndrome (RTT), a neurodevelopmental disorder that is the main cause of mental retardation in females. MeCP2 is an intrinsically disordered protein (IDP) constituted by six domains. Two domains are the main responsible elements for DNA binding (methyl-CpG binding domain, MBD) and recruitment of gene transcription\/silencing machinery (transcription repressor domain, TRD). These two domains concentrate most of the RTT-associated mutations. R106W and R133C are associated with severe and mild RTT phenotype, respectively. We have performed a comprehensive characterization of the structural and functional impact of these substitutions at molecular level. Because we have previously shown that the MBD-flanking disordered domains (N-terminal domain, NTD, and intervening domain, ID) exert a considerable influence on the structural and functional features of the MBD (Claveria-Gimeno, R. et al. Sci Rep. 2017, 7, 41635), here we report the biophysical study of the influence of the protein scaffold on the structural and functional effect induced by these two RTT-associated mutations. These results represent an example of how a given mutation may show different effects (sometimes opposing effects) depending on the molecular context.<\/jats:p>","DOI":"10.3390\/biom10111533","type":"journal-article","created":{"date-parts":[[2020,11,10]],"date-time":"2020-11-10T14:10:41Z","timestamp":1605017441000},"page":"1533","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Molecular Context-Dependent Effects Induced by Rett Syndrome-Associated Mutations in MeCP2"],"prefix":"10.3390","volume":"10","author":[{"given":"David","family":"Ortega-Alarcon","sequence":"first","affiliation":[{"name":"Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Units IQFR-CSIC-BIFI, and GBsC-CSIC-BIFI, Universidad de Zaragoza, 50018 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rafael","family":"Claveria-Gimeno","sequence":"additional","affiliation":[{"name":"Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Units IQFR-CSIC-BIFI, and GBsC-CSIC-BIFI, Universidad de Zaragoza, 50018 Zaragoza, Spain"},{"name":"Instituto Aragon\u00e9s de Ciencias de la Salud (IACS), 50009 Zaragoza, Spain"},{"name":"Instituto de Investigaci\u00f3n Sanitaria Arag\u00f3n (IIS Arag\u00f3n), 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonia","family":"Vega","sequence":"additional","affiliation":[{"name":"Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Units IQFR-CSIC-BIFI, and GBsC-CSIC-BIFI, Universidad de Zaragoza, 50018 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olga C.","family":"Jorge-Torres","sequence":"additional","affiliation":[{"name":"Josep Carreras Leukaemia Research Institute (IJC), 08916 Badalona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manel","family":"Esteller","sequence":"additional","affiliation":[{"name":"Josep Carreras Leukaemia Research Institute (IJC), 08916 Badalona, Spain"},{"name":"Centro de Investigacion Biomedica en Red Cancer (CIBERONC), 28029 Madrid, Spain"},{"name":"Institucio Catalana de Recerca i Estudis Avan\u00e7ats (ICREA), 08010 Barcelona, Spain"},{"name":"Physiological Sciences Department, School of Medicine and Health Sciences, University of Barcelona (UB), l\u2019Hospitalet de Llobregat, 08907 Barcelona, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5664-1729","authenticated-orcid":false,"given":"Olga","family":"Abian","sequence":"additional","affiliation":[{"name":"Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Units IQFR-CSIC-BIFI, and GBsC-CSIC-BIFI, Universidad de Zaragoza, 50018 Zaragoza, Spain"},{"name":"Instituto Aragon\u00e9s de Ciencias de la Salud (IACS), 50009 Zaragoza, Spain"},{"name":"Instituto de Investigaci\u00f3n Sanitaria Arag\u00f3n (IIS Arag\u00f3n), 50009 Zaragoza, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red en el \u00c1rea Tem\u00e1tica de Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd), 28029 Madrid, Spain"},{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Universidad de Zaragoza, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5702-4538","authenticated-orcid":false,"given":"Adrian","family":"Velazquez-Campoy","sequence":"additional","affiliation":[{"name":"Institute of Biocomputation and Physics of Complex Systems (BIFI), Joint Units IQFR-CSIC-BIFI, and GBsC-CSIC-BIFI, Universidad de Zaragoza, 50018 Zaragoza, Spain"},{"name":"Instituto de Investigaci\u00f3n Sanitaria Arag\u00f3n (IIS Arag\u00f3n), 50009 Zaragoza, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red en el \u00c1rea Tem\u00e1tica de Enfermedades Hep\u00e1ticas y Digestivas (CIBERehd), 28029 Madrid, Spain"},{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Universidad de Zaragoza, 50009 Zaragoza, Spain"},{"name":"Fundaci\u00f3n ARAID, Gobierno de Arag\u00f3n, 50009 Zaragoza, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,11,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.molmed.2014.03.004","article-title":"MeCP2: The long trip from a chromatin protein to neurological disorders","volume":"20","author":"Ausio","year":"2014","journal-title":"Trends Mol. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1602","DOI":"10.1016\/j.jmb.2019.10.004","article-title":"The Molecular Basis of MeCP2 Function in the Brain","volume":"432","author":"Tillotson","year":"2020","journal-title":"J. Mol. Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1139\/O10-112","article-title":"MeCP2: Structure and function","volume":"89","author":"Adkins","year":"2011","journal-title":"Biochem. Cell Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1016\/S0092-8674(00)81887-5","article-title":"MeCP2 Is a Transcriptional Repressor with Abundant Binding Sites in Genomic Chromatin","volume":"88","author":"Nan","year":"1997","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1002\/iub.386","article-title":"Binding of the Rett syndrome protein, MeCP2, to methylated and unmethylated DNA and chromatin","volume":"62","author":"Hansen","year":"2010","journal-title":"IUBMB Life"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1146\/annurev-cellbio-092910-154121","article-title":"The Role of MeCP2 in the Brain","volume":"27","author":"Guy","year":"2011","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1007\/s40142-016-0107-0","article-title":"MeCP2\u2026 Nature\u2019s wonder protein or medicine\u2019s most feared one?","volume":"4","author":"Abian","year":"2016","journal-title":"Curr. Genet. Med. Rep."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1038\/13810","article-title":"Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2","volume":"23","author":"Amir","year":"1999","journal-title":"Nat. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"253","DOI":"10.31887\/DCNS.2012.14.3\/jneul","article-title":"The relationship of Rett syndrome and MECP2 disorders to autism","volume":"14","author":"Neul","year":"2012","journal-title":"Dialog. Clin. Neurosci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1139\/O08-115","article-title":"Recent advances in MeCP2 structure and function","volume":"87","author":"Hite","year":"2009","journal-title":"Biochem. Cell Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"34115","DOI":"10.1074\/jbc.M105747200","article-title":"The Ski Protein Family Is Required for MeCP2-mediated Transcriptional Repression","volume":"276","author":"Kokura","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4886","DOI":"10.1093\/nar\/21.21.4886","article-title":"Dissection of the methyl-CpG binding domain from the chromosomal protein MeCP2","volume":"21","author":"Nan","year":"1993","journal-title":"Nucleic Acids Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1139\/O07-063","article-title":"CHD proteins: A diverse family with strong ties","volume":"85","author":"Hall","year":"2007","journal-title":"Biochem. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13073-017-0411-7","article-title":"MeCP2 mutations: Progress towards understanding and treating Rett syndrome","volume":"9","author":"Shah","year":"2017","journal-title":"Genome Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"170216","DOI":"10.1098\/rsob.170216","article-title":"Rett syndrome: A neurological disorder with metabolic components","volume":"8","author":"Kyle","year":"2018","journal-title":"Open Biol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1093\/hmg\/ddv496","article-title":"The molecular basis of variable phenotypic severity among common missense mutations causing Rett syndrome","volume":"25","author":"Brown","year":"2016","journal-title":"Hum. Mol. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1313","DOI":"10.1212\/01.wnl.0000291011.54508.aa","article-title":"Specific mutations in methyl-CpG-binding protein 2 confer different severity in Rett syndrome","volume":"70","author":"Neul","year":"2008","journal-title":"Neurology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"20523","DOI":"10.1074\/jbc.M803021200","article-title":"Rett Syndrome-causing Mutations in Human MeCP2 Result in Diverse Structural Changes That Impact Folding and DNA Interactions","volume":"283","author":"Ghosh","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4395","DOI":"10.1021\/bi9019753","article-title":"Unique Physical Properties and Interactions of the Domains of Methylated DNA Binding Protein 2","volume":"49","author":"Ghosh","year":"2010","journal-title":"Biochemistry"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"41635","DOI":"10.1038\/srep41635","article-title":"The intervening domain from MeCP2 enhances the DNA affinity of the methyl binding domain and provides an independent DNA interaction site","volume":"7","author":"Lanuza","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.ymeth.2014.09.010","article-title":"A unified framework based on the binding polynomial for characterizing biological systems by isothermal titration calorimetry","volume":"76","author":"Vega","year":"2015","journal-title":"Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/S0076-6879(08)04205-5","article-title":"Chapter 5 Isothermal Titration Calorimetry","volume":"455","author":"Freire","year":"2009","journal-title":"Methods Enzymol."},{"key":"ref_23","unstructured":"Beezer, A.E. (1980). Thermodynamics of interacting biological systems. Biological Microcalorimetry, Academic Press."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1021\/bi00784a012","article-title":"Calorimetric investigation of inhibitor binding to rabbit muscle aldolase","volume":"10","author":"Hinz","year":"1971","journal-title":"Biochemistry"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1006\/jmbi.1995.0501","article-title":"Thermodynamic Mapping of the Inhibitor Site of the Aspartic Protease Endothiapepsin","volume":"252","author":"Freire","year":"1995","journal-title":"J. Mol. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1063\/1.1416902","article-title":"Thermodynamic Quantities for the Ionization Reactions of Buffers","volume":"31","author":"Goldberg","year":"2002","journal-title":"J. Phys. Chem. Ref. Data"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1016\/S0065-3233(08)60190-4","article-title":"Linked Functions and Reciprocal Effects in Hemoglobin: A Second Look","volume":"19","author":"Wyman","year":"1964","journal-title":"Adv. Protein Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10303","DOI":"10.1021\/bi00090a005","article-title":"Circular dichroism studies of barnase and its mutants: Characterization of the contribution of aromatic side chains","volume":"32","author":"Vuilleumier","year":"1993","journal-title":"Biochemistry"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1016\/j.bbapap.2012.12.008","article-title":"Unusual biophysics of intrinsically disordered proteins","volume":"1834","author":"Uversky","year":"2013","journal-title":"Biochim. Biophys. Acta Proteins Proteom."},{"key":"ref_30","first-page":"1","article-title":"MeCP2-E1 isoform is a dynamically expressed, weakly DNA-bound protein with different protein and DNA interactions compared to MeCP2-E2","volume":"12","author":"Khajavi","year":"2019","journal-title":"Epigenetics Chromatin"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1136\/jmedgenet-2013-102113","article-title":"Methyl-CpG-binding protein 2 (MECP2) mutation type is associated with disease severity in Rett syndrome","volume":"51","author":"Cuddapah","year":"2014","journal-title":"J. Med. Genet."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/10\/11\/1533\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:31:46Z","timestamp":1760178706000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/10\/11\/1533"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,10]]},"references-count":31,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2020,11]]}},"alternative-id":["biom10111533"],"URL":"https:\/\/doi.org\/10.3390\/biom10111533","relation":{},"ISSN":["2218-273X"],"issn-type":[{"type":"electronic","value":"2218-273X"}],"subject":[],"published":{"date-parts":[[2020,11,10]]}}}