title: Single molecule visualization and characterization of Sox2-Pax6 complex formation on a regulatory DNA element using a DNA origami frame.
abstract: We report the use of atomic force microscopy (AFM) to study Sox2-Pax6 complex formation on the regulatory DNA element at a single molecule level. Using an origami DNA scaffold containing two DNA strands with different levels of tensile force, we confirmed that DNA bending is necessary for Sox2 binding. We also demonstrated that two transcription factors bind cooperatively by observing the increased occupancy of Sox2-Pax6 on the DNA element compared to that of Sox2 alone.
authors.affiliation: Department of Chemistry, Graduate School of Science, Kyoto University , Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Seigi
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Debojyoti
authors.initials: D
authors.lastname: De
authors.forename: Kumi
authors.initials: K
authors.lastname: Hidaka
authors.forename: Kyeong Kyu
authors.initials: KK
authors.lastname: Kim
authors.forename: Masayuki
authors.initials: M
authors.lastname: Endo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sugiyama
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000648:ultrastructure*; D005136:Eye Proteins / Q000737:chemistry / Q000648:ultrastructure*; D018398:Homeodomain Proteins / Q000737:chemistry / Q000648:ultrastructure*; D018625:Microscopy, Atomic Force; D046912:Multiprotein Complexes / Q000737:chemistry / Q000648:ultrastructure*; D036103:Nanotechnology / Q000379:methods; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000737:chemistry / Q000648:ultrastructure*; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry / Q000648:ultrastructure*; D055748:SOXB1 Transcription Factors / Q000737:chemistry / Q000648:ultrastructure*
chemical_list: D004268:DNA-Binding Proteins; D005136:Eye Proteins; D018398:Homeodomain Proteins; D046912:Multiprotein Complexes; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins; D055748:SOXB1 Transcription Factors; D004247:DNA