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First published online May 28, 2004
doi: 10.1242/10.1242/jcs.01131
Research Article |

1 Department of Cell Biology, National Institute for Basic Biology, Myodaiji 38, Okazaki 444-8585, Japan
2 PRESTO, Japan Science and Technology Agency, Honcho 4-1-8, Kawaguchi 332-0012, Japan
3 Department of Bio-Science, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama 526-0829, Japan
4 Department of Cell Genetics, National Institute of Genetics, Yata 1111, Mishima 411-8540, Japan
5 Department of Genetics, The Graduate University for Advanced Studies, Yata 1111, Mishima 411-8540, Japan
6 CREST, Japan Science and Technology Agency, Honcho 4-1-8, Kawaguchi 332-0012, Japan
Author for correspondence (e-mail: tamyoshi{at}lab.nig.ac.jp)
Accepted 30 January 2004
Rat LC3, a homologue of yeast Atg8 (Aut7/Apg8), localizes to autophagosomal membranes after post-translational modifications. The C-terminal fragment of LC3 is cleaved immediately following synthesis to yield a cytosolic form called LC3-I. A subpopulation of LC3-I is further converted to an autophagosome-associating form, LC3-II.
Because yeast Atg8 is conjugated with phosphatidylethanolamine (PE) by a ubiquitin-like system, it has been hypothesized that LC3 is modified in a similar manner. Here, we show that [14C]-ethanolamine was preferentially incorporated into LC3-II, suggesting that LC3-II is a PE-conjugated form. LC3-II can be a substrate of mammalian Atg4B, a homologue of yeast Atg8-PE deconjugase, supporting the idea that LC3-II is LC3-PE. Moreover, two other mammalian homologues of yeast Atg8,
-aminobutyric-acid-type-A-receptor-associated protein (GABARAP) and Golgi-associated ATPase enhancer of 16 kDa (GATE16) also generate form II, which are recovered in membrane fractions. Generation of the form II correlates with autophagosome association of GABARAP and GATE16. These results suggest that all mammalian Atg8 homologues receive a common modification to associate with autophagosomal membrane as the form II.
Key words: Autophagy, ATG, Autophagosome, UBL, Deconjugation, Phosphatidylethanolamine
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