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K. Yoshimoto, H. Hanaoka, S. Sato, T. Kato, S. Tabata, T. Noda, and Y. Ohsumi Processing of ATG8s, ubiquitin-like proteins, and their deconjugation by ATG4s are essential for plant autophagy Plant Cell 16 2004 2967 2983 The authors of this paper dissected the ATG8 conjugation step. Using a double mutant that was missing the ATG4 protease required for processing and recycling ATG8s, they showed that formation of the ATG8-PE conjugate is necessary for survival under nitrogen- and carbon-limiting growth conditions. Using ATG8-GFP fusions as reporters, punctate structures that potentially reflect autophagosomes were identified under starvation conditions by confocal fluorescence microscopy. These structures were absent in the atg4a atg4b double mutants. Possible autophagic bodies were also detected in plants treated with the vacuolar ATPase inhibitor concanamycin A. ATG8-GFP fusions might represent important markers for unequivocally visualizing autophagosomes in vivo.
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