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Exploring plant endomembrane dynamics using the photoconvertible protein Kaede.
Plant J. 2010 Aug; 63(4):696-711.PJ

Abstract

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.

Authors+Show Affiliations

Laboratoire Dynamique de la Compartimentation Cellulaire, CNRS, Institut des Sciences du Végétal, Centre de recherche de Gif (FRC3115), 91198, Gif-sur-Yvette Cedex, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20545892

Citation

Brown, Spencer C., et al. "Exploring Plant Endomembrane Dynamics Using the Photoconvertible Protein Kaede." The Plant Journal : for Cell and Molecular Biology, vol. 63, no. 4, 2010, pp. 696-711.
Brown SC, Bolte S, Gaudin M, et al. Exploring plant endomembrane dynamics using the photoconvertible protein Kaede. Plant J. 2010;63(4):696-711.
Brown, S. C., Bolte, S., Gaudin, M., Pereira, C., Marion, J., Soler, M. N., & Satiat-Jeunemaitre, B. (2010). Exploring plant endomembrane dynamics using the photoconvertible protein Kaede. The Plant Journal : for Cell and Molecular Biology, 63(4), 696-711. https://doi.org/10.1111/j.1365-313X.2010.04272.x
Brown SC, et al. Exploring Plant Endomembrane Dynamics Using the Photoconvertible Protein Kaede. Plant J. 2010;63(4):696-711. PubMed PMID: 20545892.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Exploring plant endomembrane dynamics using the photoconvertible protein Kaede. AU - Brown,Spencer C, AU - Bolte,Susanne, AU - Gaudin,Marie, AU - Pereira,Claudia, AU - Marion,Jessica, AU - Soler,Marie-Noëlle, AU - Satiat-Jeunemaitre,Béatrice, PY - 2010/6/16/entrez PY - 2010/6/16/pubmed PY - 2011/3/22/medline SP - 696 EP - 711 JF - The Plant journal : for cell and molecular biology JO - Plant J. VL - 63 IS - 4 N2 - 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. SN - 1365-313X UR - https://www.unboundmedicine.com/medline/citation/20545892/Exploring_plant_endomembrane_dynamics_using_the_photoconvertible_protein_Kaede_ L2 - https://doi.org/10.1111/j.1365-313X.2010.04272.x DB - PRIME DP - Unbound Medicine ER -