title: In-depth Characterization of the Homodimerization Domain of the Transcription Factor THAP1 and Dystonia-Causing Mutations Therein.
abstract: Mutations in the THAP1 gene encoding the transcription factor THAP1 have been shown to cause DYT6 dystonia. THAP1 contains a highly conserved THAP zinc finger at its N-terminal region which allows specific binding to its target sequences as well as a coiled-coil domain (amino acids 139-190) towards its C-terminus postulated as a protein-protein-binding motif. While several DYT6-causing mutations within the THAP domain were shown to decrease THAP1 activity in transcriptional regulation and DNA-binding, the role of mutations within the coiled-coil domain is rather unknown. Therefore, assigning a function to this domain may enable functional testing of mutations in this region. Notably, THAP1 and other THAP proteins form homodimers; however, the responsible domain has not been elucidated in detail. We show that the region of amino acids 139-185 is involved in formation of THAP1 homodimers by using yeast-two-hybrid, GST pull-down, and cross-linking assays. Surprisingly, all nine reported DYT6-causing missense mutations within this region had no effect on dimerization of THAP1 in GST pull-down and formaldehyde cross-linking assays. In conclusion, we demonstrated that a region of 47 amino acids is involved in THAP1 homodimerization but mutations in this region seem not to impair this mechanism.
authors.affiliation: Sektion für Funktionelle Genetik am Institut für Humangenetik, Universität zu Lübeck, 23538, Luebeck, Germany.
authors.forename: Alev
authors.initials: A
authors.lastname: Richter
authors.affiliation: Sektion für Funktionelle Genetik am Institut für Humangenetik, Universität zu Lübeck, 23538, Luebeck, Germany.
authors.forename: Ronja
authors.initials: R
authors.lastname: Hollstein
authors.affiliation: Institute of Neurogenetics, University of Luebeck, Ratzeburger Allee 160, 23538, Luebeck, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Hebert
authors.affiliation: Institute of Neurogenetics, University of Luebeck, Ratzeburger Allee 160, 23538, Luebeck, Germany.
authors.forename: Franca
authors.initials: F
authors.lastname: Vulinovic
authors.affiliation: Sektion für Funktionelle Genetik am Institut für Humangenetik, Universität zu Lübeck, 23538, Luebeck, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Eckhold
authors.affiliation: Sektion für Funktionelle Genetik am Institut für Humangenetik, Universität zu Lübeck, 23538, Luebeck, Germany.
authors.forename: Alma
authors.initials: A
authors.lastname: Osmanovic
authors.affiliation: Institute of Physiology, Center for Structural and Cell Biology in Medicine, University of Luebeck, 23538, Luebeck, Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Depping
authors.affiliation: Sektion für Funktionelle Genetik am Institut für Humangenetik, Universität zu Lübeck, 23538, Luebeck, Germany.
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Kaiser
authors.affiliation: Institute of Neurogenetics, University of Luebeck, Ratzeburger Allee 160, 23538, Luebeck, Germany. katja.lohmann@neuro.uni-luebeck.de.
authors.forename: Katja
authors.initials: K
authors.lastname: Lohmann
mesh_terms: D051017:Apoptosis Regulatory Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004421:Dystonia / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000072417:Protein Domains; D055503:Protein Multimerization
chemical_list: D051017:Apoptosis Regulatory Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C474080:THAP1 protein, human
keywords: Dystonia; Homodimerization; Mutation; Transcription