title: Multimerization of the mouse TATA-binding protein (TBP) driven by its C-terminal conserved domain.
abstract: The conformational states of the mouse TATA-binding protein (TBP) in solution were studied. A histidine tag and a factor Xa recognition site-carrying mouse TBP was expressed in E. coli, highly purified, and its fundamental functions as a TBP were demonstrated. We analyzed the molecular states of mouse TBP by gel filtration and glycerol gradient sedimentation, and found that TBP forms heterogeneous multimers in solution. Direct binding of TBP molecules to each other was proven by the far-Western procedure. Analyses using TBPs truncated at the N- and C-termini demonstrated that the functionally important C-terminal domain was responsible for homomultimer formation, and the N-terminal domain enhances multimerization. Furthermore, it was found that the TATA sequence dissociates homomultimers, and only monomeric TBP binds to the TATA-box. We suggest that TBP shares structural motifs in the C-terminal conserved domain for intermolecular interaction and TATA-binding.
authors.affiliation: Department of Biology, Faculty of Science, Chiba University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kato
authors.forename: Y
authors.initials: Y
authors.lastname: Makino
authors.forename: T
authors.initials: T
authors.lastname: Kishimoto
authors.forename: J
authors.initials: J
authors.lastname: Yamauchi
authors.forename: S
authors.initials: S
authors.lastname: Kato
authors.forename: M
authors.initials: M
authors.lastname: Muramatsu
authors.forename: T
authors.initials: T
authors.lastname: Tamura
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017124:Conserved Sequence; D004247:DNA; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D015951:Factor Xa / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012996:Solutions; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D012996:Solutions; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; D004247:DNA; D015951:Factor Xa