Observation of the Yarrowia lipolytica peroxisome-vacuole dynamics by fluorescence microscopy with a single filter set.
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.
Nazarko
Taras Y
TY
Institute of Cell Biology, National Academy of Sciences of Ukraine, Drahomanov Street, 14/16, 79005 Lviv, Ukraine.
Nicaud
Jean-Marc
JM
Sibirny
Andrei A
AA
D042962:Acyl-CoA Oxidase; D001426:Bacterial Proteins; D005021:Ethylamines / Q000494:pharmacology; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence / Q000379:methods*; D019301:Oleic Acid / Q000494:pharmacology; D020675:Peroxisomes / Q000187:drug effects / Q000502:physiology*; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D011993:Recombinant Fusion Proteins; D014617:Vacuoles / Q000502:physiology*; D025062:Yarrowia / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*
D001426:Bacterial Proteins; D005021:Ethylamines; C092350:FM 4-64; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D019301:Oleic Acid; D042962:Acyl-CoA Oxidase