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Visualizing Dynamics of Cell Signaling In Vivo with a Phase Separation-Based Kinase Reporter.
Zhang, Qiang; Huang, Hai; Zhang, Luqing; Wu, Roland; Chung, Chan-I; Zhang, Shao-Qing; Torra, Joaquim; Schepis, Antonino; Coughlin, Shaun R; Kornberg, Thomas B; Shu, Xiaokun.
  • Zhang Q; Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Huang H; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Zhang L; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Wu R; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Chung CI; Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Zhang SQ; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Torra J; Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA; Institut Químic de Sarrià, Universitat Ramon Llull, Via Augusta 390, 08017 Barcelon
  • Schepis A; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Coughlin SR; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Kornberg TB; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA.
  • Shu X; Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94143, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94143, USA. Electronic address: xiaokun.shu@ucsf.edu.
Mol Cell ; 69(2): 334-346.e4, 2018 01 18.
Article en En | MEDLINE | ID: mdl-29307513
ABSTRACT
Visualizing dynamics of kinase activity in living animals is essential for mechanistic understanding of cell and developmental biology. We describe GFP-based kinase reporters that phase-separate upon kinase activation via multivalent protein-protein interactions, forming intensively fluorescent droplets. Called SPARK (separation of phases-based activity reporter of kinase), these reporters have large dynamic range (fluorescence change), high brightness, fast kinetics, and are reversible. The SPARK-based protein kinase A (PKA) reporter reveals oscillatory dynamics of PKA activities upon G protein-coupled receptor activation. The SPARK-based extracellular signal-regulated kinase (ERK) reporter unveils transient dynamics of ERK activity during tracheal metamorphosis in live Drosophila. Because of intensive brightness and simple signal pattern, SPARKs allow easy examination of kinase signaling in living animals in a qualitative way. The modular design of SPARK will facilitate development of reporters of other kinases.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Fosfotransferasas / Transducción de Señal / Imagen Óptica Tipo de estudio: Qualitative_research Límite: Animals / Humans Idioma: En Año: 2018 Tipo del documento: Article
Texto completo
Imprimir
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PubMed Links
Search on Google

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Fosfotransferasas / Transducción de Señal / Imagen Óptica Tipo de estudio: Qualitative_research Límite: Animals / Humans Idioma: En Año: 2018 Tipo del documento: Article