title: Synthetic protein-protein interaction domains created by shuffling Cys2His2 zinc-fingers.
abstract: Cys2His2 zinc-fingers (C2H2 ZFs) mediate a wide variety of protein-DNA and protein-protein interactions. DNA-binding C2H2 ZFs can be shuffled to yield artificial proteins with different DNA binding specificities. Here we demonstrate that shuffling of C2H2 ZFs from transcription factor dimerization zinc-finger (DZF) domains can also yield two-finger DZFs with novel protein-protein interaction specificities. We show that these synthetic protein-protein interaction domains can be used to mediate activation of a single-copy reporter gene in bacterial cells and of an endogenous gene in human cells. In addition, the synthetic two-finger domains we constructed can also be linked together to create more extended, four-finger interfaces. Our results demonstrate that shuffling of C2H2 ZFs can yield artificial protein-interaction components that should be useful for applications in synthetic biology.
authors.affiliation: Molecular Pathology Unit, Department of Pathology, Massachusetts General Hospital, Charlestown, MA 02129, USA.
authors.forename: Astrid V
authors.initials: AV
authors.lastname: Giesecke
authors.forename: Rui
authors.initials: R
authors.lastname: Fang
authors.forename: J Keith
authors.initials: JK
authors.lastname: Joung
mesh_terms: D001665:Binding Sites; D003545:Cysteine; D004268:DNA-Binding Proteins / Q000138:chemical synthesis; D017930:Genes, Reporter; D006639:Histidine; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D016335:Zinc Fingers
chemical_list: D004268:DNA-Binding Proteins; D006639:Histidine; D003545:Cysteine