Exploring plant endomembrane dynamics using the photoconvertible protein Kaede

• Tipo: Artigo em Revista Científica Internacional
• Data: 2010

Autores

Brown, SC (Autor) Outra; Bolte, S (Autor) Outra; Gaudin, M (Autor) Outra; Claudia Pereira (Autor) Outra; Marion, J (Autor) Outra; Soler, MN (Autor) Outra; Satiat Jeunemaitre, B (Autor) Outra

Revista

Título: Plant Journal

Vol. 63

Páginas: 696-711

ISSN: 0960-7412

Editora: Wiley-Blackwell

Indexação

ISI Web of Knowledge - 23 Citações

Scopus - 16 Citações

Outras Informações

ID Authenticus: P-003-4JS

DOI: 10.1111/j.1365-313x.2010.04272.x

Abstract (EN): P>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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 16