title: Structure-based design of a potent artificial transactivation domain based on p53.
abstract: Malfunctions in transcriptional regulation are associated with a number of critical human diseases. As a result, there is considerable interest in designing artificial transcription activators (ATAs) that specifically control genes linked to human diseases. Like native transcriptional activator proteins, an ATA must minimally contain a DNA-binding domain (DBD) and a transactivation domain (TAD) and, although there are several reliable methods for designing artificial DBDs, designing artificial TADs has proven difficult. In this manuscript, we present a structure-based strategy for designing short peptides containing natural amino acids that function as artificial TADs. Using a segment of the TAD of p53 as the scaffolding, modifications are introduced to increase the helical propensity of the peptides. The most active artificial TAD, termed E-Cap-(LL), is a 13-mer peptide that contains four key residues from p53, an N-capping motif and a dileucine hydrophobic bridge. In vitro analysis demonstrates that E-Cap-(LL) interacts with several known p53 target proteins, while in vivo studies in a yeast model system show that it is a 20-fold more potent transcriptional activator than the native p53-13 peptide. These results demonstrate that structure-based design represents a promising approach for developing artificial TADs that can be combined with artificial DBDs to create potent and specific ATAs.
authors.affiliation: Département de Biochimie, Université de Montréal, C.P. 6128 Succursale, Centre-Ville, Montréal, Quebec H3C 3J7, Canada.
authors.forename: Chantal
authors.initials: C
authors.lastname: Langlois
authors.forename: Annarita
authors.initials: A
authors.lastname: Del Gatto
authors.forename: Geneviève
authors.initials: G
authors.lastname: Arseneault
authors.forename: Julien
authors.initials: J
authors.lastname: Lafrance-Vanasse
authors.forename: Mariarosaria
authors.initials: M
authors.lastname: De Simone
authors.forename: Thomas
authors.initials: T
authors.lastname: Morse
authors.forename: Ivan
authors.initials: I
authors.lastname: de Paola
authors.forename: Mathieu
authors.initials: M
authors.lastname: Lussier-Price
authors.forename: Pascale
authors.initials: P
authors.lastname: Legault
authors.forename: Carlo
authors.initials: C
authors.lastname: Pedone
authors.forename: Laura
authors.initials: L
authors.lastname: Zaccaro
authors.forename: James G
authors.initials: JG
authors.lastname: Omichinski
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D007930:Leucine / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics
chemical_list: D010455:Peptides; D016159:Tumor Suppressor Protein p53; D007930:Leucine