title: Interaction of the transactivation domain of B-Myb with the TAZ2 domain of the coactivator p300: molecular features and properties of the complex.
abstract: The transcription factor B-Myb is a key regulator of the cell cycle in vertebrates, with activation of transcription involving the recognition of specific DNA target sites and the recruitment of functional partner proteins, including the coactivators p300 and CBP. Here we report the results of detailed studies of the interaction between the transactivation domain of B-Myb (B-Myb TAD) and the TAZ2 domain of p300. The B-Myb TAD was characterized using circular dichroism, fluorescence and NMR spectroscopy, which revealed that the isolated domain exists as a random coil polypeptide. Pull-down and spectroscopic experiments clearly showed that the B-Myb TAD binds to p300 TAZ2 to form a moderately tight (K(d) ~1.0-10 µM) complex, which results in at least partial folding of the B-Myb TAD. Significant changes in NMR spectra of p300 TAZ2 suggest that the B-Myb TAD binds to a relatively large patch on the surface of the domain (~1200 Å(2)). The apparent B-Myb TAD binding site on p300 TAZ2 shows striking similarity to the surface of CBP TAZ2 involved in binding to the transactivation domain of the transcription factor signal transducer and activator of transcription 1 (STAT1), which suggests that the structure of the B-Myb TAD-p300 TAZ2 complex may share many features with that reported for STAT1 TAD-p300 TAZ2.
authors.affiliation: Department of Biochemistry, University of Leicester, Henry Wellcome Building, Leicester, United Kingdom.
authors.forename: Ojore
authors.initials: O
authors.lastname: Oka
authors.forename: Lorna C
authors.initials: LC
authors.lastname: Waters
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Strong
authors.forename: Nuvjeevan S
authors.initials: NS
authors.lastname: Dosanjh
authors.forename: Vaclav
authors.initials: V
authors.lastname: Veverka
authors.forename: Frederick W
authors.initials: FW
authors.lastname: Muskett
authors.forename: Philip S
authors.initials: PS
authors.lastname: Renshaw
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Klempnauer
authors.forename: Mark D
authors.initials: MD
authors.lastname: Carr
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism*; D002942:Circular Dichroism; D050881:E1A-Associated p300 Protein / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D015534:Trans-Activators / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; C064352:MYBL2 protein, human; D015534:Trans-Activators; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human