Exploring plant endomembrane dynamics using the photoconvertible protein Kaede

Authors

  • Spencer C. Brown,

    1. Laboratoire Dynamique de la Compartimentation Cellulaire, CNRS, Institut des Sciences du Végétal, Centre de recherche de Gif (FRC3115), 91198, Gif-sur-Yvette Cedex, France
    Search for more papers by this author
  • Susanne Bolte,

    1. Plateforme de Microscopie Photonique, Pôle de Biologie Cellulaire, Imagif, Centre de Recherche de Gif, (FRC3115), CNRS, IFR 87, 91198, Gif-sur-Yvette Cedex, France
    Search for more papers by this author
  • Marie Gaudin,

    1. Laboratoire Dynamique de la Compartimentation Cellulaire, CNRS, Institut des Sciences du Végétal, Centre de recherche de Gif (FRC3115), 91198, Gif-sur-Yvette Cedex, France
    Search for more papers by this author
  • Claudia Pereira,

    1. Faculdade de Ciências da Universidade do Porto, Departamento de Botânica, Rua do Campo Alegre, s/n, 4169-007, Porto, Portugal
    Search for more papers by this author
  • Jessica Marion,

    1. Laboratoire Dynamique de la Compartimentation Cellulaire, CNRS, Institut des Sciences du Végétal, Centre de recherche de Gif (FRC3115), 91198, Gif-sur-Yvette Cedex, France
    Search for more papers by this author
  • Marie-Noëlle Soler,

    1. Plateforme de Microscopie Photonique, Pôle de Biologie Cellulaire, Imagif, Centre de Recherche de Gif, (FRC3115), CNRS, IFR 87, 91198, Gif-sur-Yvette Cedex, France
    Search for more papers by this author
  • Béatrice Satiat-Jeunemaitre

    Corresponding author
    1. Laboratoire Dynamique de la Compartimentation Cellulaire, CNRS, Institut des Sciences du Végétal, Centre de recherche de Gif (FRC3115), 91198, Gif-sur-Yvette Cedex, France
    2. Plateforme de Microscopie Photonique, Pôle de Biologie Cellulaire, Imagif, Centre de Recherche de Gif, (FRC3115), CNRS, IFR 87, 91198, Gif-sur-Yvette Cedex, France
    Search for more papers by this author

For correspondence (fax +33 0 1 69823695; e-mail bsj@isv.cnrs-gif.fr).

Summary

Photoactivatable and photoconvertible fluorescent proteins capable of pronounced light-induced spectral changes are a powerful addition to the fluorescent protein toolbox of the cell biologist. They permit specific tracking of one subcellular structure (organelle or cell subdomain) within a differentially labelled population. They also enable pulse–chase analysis of protein traffic. The Kaede gene codes for a tetrameric protein found in the stony coral Trachyphyllia geoffroyi, which emits green fluorescence that irreversibly shifts to red following radiation with UV or violet light. We report here the use of Kaede to explore the plant secretory pathway. Kaede versions of the Golgi marker sialyl-transferase (ST-Kaede) and of the vacuolar pathway marker cardosin A (cardA-Kaede) were engineered. Several optical devices enabling photoconversion and observation of Kaede using these two constructs were assessed to optimize Kaede-based imaging protocols. Photoconverted ST-Kaede red-labelled organelles can be followed within neighbouring populations of non-converted green Golgi stacks, by their gradual development of orange/yellow coloration from de novo synthesis of Golgi proteins (green). Results highlight some aspects on the dynamics of the plant Golgi. For plant bio-imaging, the photoconvertible Kaede offers a powerful tool to track the dynamic behaviour of designated subpopulations of Golgi within living cells, while visualizing the de novo formation of proteins and structures, such as a Golgi stack.

Ancillary