title: Subunit interaction in the CCAAT-binding heteromeric complex is mediated by a very short alpha-helix in HAP2.
abstract: We dissected the domain of HAP2 that mediates subunit association in the heteromeric CCAAT-binding complex, first by genetic mutational analysis and then by structural studies. The mutational data suggest that a very short region in HAP2 mediates protein-protein association and that the structure of this domain is likely to be an alpha-helix. The CD analyses of a 15-residue synthetic oligopeptide covering this region confirm this surmise. The oligopeptide indeed formed an unusually thermal stable alpha-helix in aqueous solution. Eight amino acids that lie along one face of this helix, including three arginines, are found to be critical for protein-protein association. The partner that interacts with this helical motif is likely to be another subunit in the HAP complex, since the CCAAT-binding factor is shown to contain one molecule of HAP2. Our results suggest that very short regions in proteins can encode precise structures and mediate stable and specific protein-protein recognition and interactions.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge 02139-4307.
authors.forename: Y
authors.initials: Y
authors.lastname: Xing
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.forename: J T
authors.initials: JT
authors.lastname: Olesen
authors.forename: A
authors.initials: A
authors.lastname: Rich
authors.forename: L
authors.initials: L
authors.lastname: Guarente
mesh_terms: D000595:Amino Acid Sequence; D023081:CCAAT-Binding Factor; D002942:Circular Dichroism; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism; D016350:Leucine Zippers; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D023081:CCAAT-Binding Factor; D005656:Fungal Proteins; D046911:Macromolecular Substances; D014157:Transcription Factors