title: Single-Molecule Imaging Reveals Dynamics of CREB Transcription Factor Bound to Its Target Sequence.
abstract: Proper spatiotemporal gene expression is achieved by selective DNA binding of transcription factors in the genome. The most intriguing question is how dynamic interactions between transcription factors and their target sites contribute to gene regulation by recruiting the basal transcriptional machinery. Here we demonstrate individual binding and dissociation events of the transcription factor cAMP response element-binding protein (CREB), both in vitro and in living cells, using single-molecule imaging. Fluorescent-tagged CREB bound to its target sequence cAMP-response element (CRE) for a remarkably longer period (dissociation rate constant: 0.21 s(-1)) than to an unrelated sequence (2.74 s(-1)). Moreover, CREB resided at restricted positions in the living cell nucleus for a comparable period. These results suggest that CREB stimulates transcription by binding transiently to CRE in the time range of several seconds.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Sugo
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Morimatsu
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Arai
authors.affiliation: Graduate School of Engineering Science, Osaka University, Osaka, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kousoku
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Ohkuni
authors.affiliation: Graduate School of Engineering Science, Osaka University, Osaka, Japan.
authors.forename: Taishin
authors.initials: T
authors.lastname: Nomura
authors.affiliation: 1] Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan [2] Riken Quantitative Biological Center (QBic), Osaka, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Yanagida
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D057054:Molecular Imaging / Q000379:methods; D011485:Protein Binding; D020218:Response Elements
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein