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In cases where the mutant gene encodes a transcription factor, the dominant-negative mode of action of the mutant protein is particularly poorly understood. Here, we studied the genome-wide mechanism underlying a dominant-negative form of the SOX18 transcription factor (SOX18RaOp) responsible for both the classical mouse mutant Ragged Opossum and the human genetic disorder Hypotrichosis-lymphedema-telangiectasia-renal defect syndrome. Combining three single-molecule imaging assays in living cells together with genomics and proteomics analysis, we found that SOX18RaOp disrupts the system through an accumulation of molecular interferences which impair several functional properties of the wild-type SOX18 protein, including its target gene selection process. The dominant-negative effect is further amplified by poisoning the interactome of its wild-type counterpart, which perturbs regulatory nodes such as SOX7 and MEF2C. Our findings explain in unprecedented detail the multi-layered process that underpins the molecular aetiology of dominant-negative transcription factor function.<\/jats:p>","DOI":"10.1093\/nar\/gkab820","type":"journal-article","created":{"date-parts":[[2021,9,8]],"date-time":"2021-09-08T07:37:07Z","timestamp":1631086627000},"page":"10931-10955","source":"Crossref","is-referenced-by-count":23,"title":["A dominant-negative SOX18 mutant disrupts multiple regulatory layers essential to transcription factor activity"],"prefix":"10.1093","volume":"49","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2984-5782","authenticated-orcid":false,"given":"Alex J","family":"McCann","sequence":"first","affiliation":[{"name":"Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD\u00a04072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jieqiong","family":"Lou","sequence":"additional","affiliation":[{"name":"School of Physics, Department of Biochemistry and Molecular Biology, Bio21, The University of Melbourne, Melbourne, VIC\u00a03010, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehdi","family":"Moustaqil","sequence":"additional","affiliation":[{"name":"EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW\u00a01466, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew S","family":"Graus","sequence":"additional","affiliation":[{"name":"The David Richmond Laboratory for Cardio-Vascular Development: gene regulation and editing, The Centenary Institute, Newtown, Sydney, NSW\u00a02006, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ailisa","family":"Blum","sequence":"additional","affiliation":[{"name":"Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, QLD\u00a04072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frank","family":"Fontaine","sequence":"additional","affiliation":[{"name":"Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD\u00a04072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Liu","sequence":"additional","affiliation":[{"name":"Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA\u00a020147, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Winnie","family":"Luu","sequence":"additional","affiliation":[{"name":"The David Richmond Laboratory for Cardio-Vascular Development: gene regulation and editing, The Centenary Institute, Newtown, Sydney, NSW\u00a02006, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulina","family":"Rudolffi-Soto","sequence":"additional","affiliation":[{"name":"EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW\u00a01466, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Koopman","sequence":"additional","affiliation":[{"name":"Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD\u00a04072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Sierecki","sequence":"additional","affiliation":[{"name":"EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW\u00a01466, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yann","family":"Gambin","sequence":"additional","affiliation":[{"name":"EMBL Australia Node in Single Molecule Science and School of Medical Sciences, The University of New South Wales, Sydney, NSW\u00a01466, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fr\u00e9d\u00e9ric\u00a0A","family":"Meunier","sequence":"additional","affiliation":[{"name":"Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, QLD\u00a04072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhe","family":"Liu","sequence":"additional","affiliation":[{"name":"Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA\u00a020147, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth","family":"Hinde","sequence":"additional","affiliation":[{"name":"School of Physics, Department of Biochemistry and Molecular Biology, Bio21, The University of Melbourne, Melbourne, VIC\u00a03010, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathias","family":"Francois","sequence":"additional","affiliation":[{"name":"Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD\u00a04072, Australia"},{"name":"The David Richmond Laboratory for Cardio-Vascular Development: gene regulation and editing, The Centenary Institute, Newtown, Sydney, NSW\u00a02006, Australia"},{"name":"School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW\u00a02006, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,9,27]]},"reference":[{"key":"2021110312470301700_B1","doi-asserted-by":"crossref","first-page":"e04236","DOI":"10.7554\/eLife.04236","article-title":"3D imaging of Sox2 enhancer clusters in embryonic stem cells","volume":"3","author":"Liu","year":"2014","journal-title":"Elife"},{"key":"2021110312470301700_B2","doi-asserted-by":"crossref","first-page":"1274","DOI":"10.1016\/j.cell.2014.01.062","article-title":"Single-molecule dynamics of enhanceosome assembly in embryonic stem 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