title: Positive and negative cooperativity of modularly assembled zinc fingers.
abstract: One simple and widespread method to create engineered zinc fingers targeting the desired DNA sequences is to modularly assemble multiple finger modules pre-selected to recognize each DNA triplet. However, it has become known that a sufficient DNA binding affinity is not always obtained. In order to create successful zinc finger proteins, it is important to understand the context-dependent contribution of each finger module to the DNA binding ability of the assembled zinc finger proteins. Here, we have created finger-deletion mutants of zinc finger proteins and examined the DNA bindings of these zinc fingers to clarify the contributions of each finger module. Our results indicate that not only a positive cooperativity but also a context-dependent reduction in the DNA binding activity can be induced by assembling zinc finger modules.
authors.affiliation: Institute for Chemical Research, Kyoto University, Uji, Kyoto, Japan. imiki@scl.kyoto-u.ac.jp
authors.forename: Miki
authors.initials: M
authors.lastname: Imanishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nakamura
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Morisaki
authors.forename: Shiroh
authors.initials: S
authors.lastname: Futaki
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D024721:E-Box Elements; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D051379:Mice; D056950:Period Circadian Proteins; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014158:Transcription, Genetic; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C539023:Per1 protein, mouse; D056950:Period Circadian Proteins; D004247:DNA