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Genetically Encoded Tools for Optical Dissection of the Mammalian Cell Cycle.
Sakaue-Sawano, Asako; Yo, Masahiro; Komatsu, Naoki; Hiratsuka, Toru; Kogure, Takako; Hoshida, Tetsushi; Goshima, Naoki; Matsuda, Michiyuki; Miyoshi, Hiroyuki; Miyawaki, Atsushi.
Affiliation
  • Sakaue-Sawano A; Laboratory for Cell Function Dynamics, BSI, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
  • Yo M; Laboratory for Cell Function Dynamics, BSI, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
  • Komatsu N; Laboratory for Cell Function Dynamics, BSI, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
  • Hiratsuka T; Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan; King's College London Centre for Stem Cells and Regenerative Medicine, 28th Floor, Tower Wing, Guy's Campus, Great Maze Pond, London SE1 9RT, UK.
  • Kogure T; Laboratory for Cell Function Dynamics, BSI, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
  • Hoshida T; Biotechnological Optics Research Team, Center for Advanced Photonics, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.
  • Goshima N; Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Koutou 135-0064, Japan.
  • Matsuda M; Department of Pathology and Biology of Diseases, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
  • Miyoshi H; Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
  • Miyawaki A; Laboratory for Cell Function Dynamics, BSI, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan; Biotechnological Optics Research Team, Center for Advanced Photonics, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan. Electronic address: matsushi@brain.riken.jp.
Mol Cell ; 68(3): 626-640.e5, 2017 11 02.
Article in En | MEDLINE | ID: mdl-29107535
ABSTRACT
Eukaryotic cells spend most of their life in interphase of the cell cycle. Understanding the rich diversity of metabolic and genomic regulation that occurs in interphase requires the demarcation of precise phase boundaries in situ. Here, we report the properties of two genetically encoded fluorescence sensors, Fucci(CA) and Fucci(SCA), which enable real-time monitoring of interphase and cell-cycle biology. We re-engineered the Cdt1-based sensor from the original Fucci system to respond to S phase-specific CUL4Ddb1-mediated ubiquitylation alone or in combination with SCFSkp2-mediated ubiquitylation. In cultured cells, Fucci(CA) produced a sharp triple color-distinct separation of G1, S, and G2, while Fucci(SCA) permitted a two-color readout of G1 and S/G2. Fucci(CA) applications included tracking the transient G1 phase of rapidly dividing mouse embryonic stem cells and identifying a window for UV-irradiation damage in S phase. These results show that Fucci(CA) is an essential tool for quantitative studies of interphase cell-cycle regulation.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Cell Cycle / Cell Cycle Proteins / Cullin Proteins / Embryonic Stem Cells / Fluorescent Dyes / Luminescent Proteins Limits: Animals / Humans Language: En Journal: Mol Cell Journal subject: BIOLOGIA MOLECULAR Year: 2017 Type: Article Affiliation country: Japan
Fulltext
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Cell Cycle / Cell Cycle Proteins / Cullin Proteins / Embryonic Stem Cells / Fluorescent Dyes / Luminescent Proteins Limits: Animals / Humans Language: En Journal: Mol Cell Journal subject: BIOLOGIA MOLECULAR Year: 2017 Type: Article Affiliation country: Japan