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Observation of the Yarrowia lipolytica peroxisome—vacuole dynamics by fluorescence microscopy with a single filter set
Author(s) -
Nazarko Taras Y.,
Nicaud JeanMarc,
Sibirny Andrei A.
Publication year - 2005
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2004.11.014
Subject(s) - yarrowia , peroxisome , vacuole , microbody , biochemistry , yeast , biology , yellow fluorescent protein , fluorescence , fluorescein isothiocyanate , chemistry , cytoplasm , gene , physics , quantum mechanics
We constructed the fusion of peroxisomal acyl‐CoA oxidase 3 and the enhanced yellow fluorescent protein (EYFP) for fluorescent labeling of Yarrowia lipolytica peroxisomes. Using the spectral overlap between EYFP and FM4‐64, we developed a procedure for simultaneous observation of Y. lipolytica peroxisomes and vacuoles with the single fluorescein isothiocyanate filter set. Using this procedure we were able to follow the Y. lipolytica peroxisome—vacuole dynamics under pexophagy conditions and show that Y. lipolytica peroxisomes are degraded in the vacuoles by a macropexophagic mechanism.