Premium
A simple estimation of peroxisomal degradation with green fluorescent protein – an application for cell cycle analysis
Author(s) -
Arai Kunizo,
Ohkuma Shoji,
Matsukawa Toru,
Kato Satoru
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02964-7
Subject(s) - peroxisome , green fluorescent protein , cytosol , organelle , microbiology and biotechnology , autophagy , fluorescence , protein degradation , biochemistry , biology , chemistry , enzyme , gene , apoptosis , physics , quantum mechanics
When nutrients are depleted from the environment, mammalian cells begin to degrade their own cytosol and organelles. This bulk protein degradation is mediated by autophagy. In this study, peroxisomes in living CHO‐K1 cells were visualized by targeting the green fluorescent protein (GFP) tagged with a type 1 peroxisomal targeting signal. The nutrient‐starved condition induced a decay of GFP fluorescence in the peroxisomes and autophagic inhibitors such as 3‐methyladenine suppressed the decay of GFP fluorescence (13–60% of starvation). By double labeling the nuclear DNA and peroxisomal GFP, the autophagy specifically occurred at the G1 phase of the cell cycle and the autophagic inhibitors suppressed the G1 arrest. The vital stain technique with GFP is a very simple and useful marker to quantitatively estimate or to further study peroxisomal degradation.