Positive and negative cooperativity of modularly assembled zinc fingers.
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.
Imanishi
Miki
M
Institute for Chemical Research, Kyoto University, Uji, Kyoto, Japan. imiki@scl.kyoto-u.ac.jp
Nakamura
Atsushi
A
Morisaki
Tatsuya
T
Futaki
Shiroh
S
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*
D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C539023:Per1 protein, mouse; D056950:Period Circadian Proteins; D004247:DNA