title: Three-Dimensional, Live-Cell Imaging of Chromatin Dynamics in Plant Nuclei Using Chromatin Tagging Systems.
abstract: In plants, chromatin dynamics spatiotemporally change in response to various environmental stimuli. However, little is known about chromatin dynamics in the nuclei of plants. Here, we introduce a three-dimensional, live-cell imaging method that can monitor chromatin dynamics in nuclei via a chromatin tagging system that can visualize specific genomic loci in living plant cells. The chromatin tagging system is based on a bacterial operator/repressor system in which the repressor is fused to fluorescent proteins. A recent refinement of promoters for the system solved the problem of gene silencing and abnormal pairing frequencies between operators. Using this system, we can detect the spatiotemporal dynamics of two homologous loci as two fluorescent signals within a nucleus and monitor the distance between homologous loci. These live-cell imaging methods will provide new insights into genome organization, development processes, and subnuclear responses to environmental stimuli in plants.
authors.affiliation: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641, Yamazaki, Noda, Chiba, 278-8510, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Hirakawa
authors.affiliation: Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641, Yamazaki, Noda, Chiba, 278-8510, Japan. sachi@rs.tus.ac.jp.
authors.forename: Sachihiro
authors.initials: S
authors.lastname: Matsunaga
mesh_terms: D017360:Arabidopsis / Q000235:genetics; D002467:Cell Nucleus / Q000235:genetics*; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D018548:Cotyledon / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D056968:Lac Repressors / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics
chemical_list: D002843:Chromatin; D056968:Lac Repressors; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Chromatin dynamics; Chromatin tagging system; Live-cell imaging; Time-lapse imaging; lacO/LacI-EGFP