tailieunhanh - Báo cáo khoa học: Atg8L/Apg8L is the fourth mammalian modifier of mammalian Atg8 conjugation mediated by human Atg4B, Atg7 and Atg3

Murine Atg8L⁄Apg8L has significant homology with the other known mammalian Atg8 homologs, LC3, GABARAP and GATE-16. However, it is unclear whether murine Atg8L modification is mediated by human Atg4B, Atg7 and Atg3. Expression of Atg8L in HEK293 cells led to clea-vage of its vitro, the C-terminus of Atg8L was cleaved by human Atg4B, but not human Atg4A or Atg4C. | ềFEBS Journal Atg8L Apg8L is the fourth mammalian modifier of mammalian Atg8 conjugation mediated by human Atg4B Atg7 and Atg3 Isei Tanida Yu-shin Sou Naoko Minematsu-Ikeguchi Takashi Ueno and Eiki Kominami Molecular CellBiology Department of Biochemistry Juntendo University Schoolof Medicine Tokyo Japan Keywords autophagy GABARAP GATE-16 LC3 ubiquitylation-like modification Correspondence E. Kominami Department of Biochemistry Juntendo University Schoolof Medicine 2-1-1 Hongo Bunkyo-ku Tokyo 113-8421 Japan Fax 81 3 5802 5889 Tel 81 3 5802 1031 E-mail kominami@ Received 23 December 2005 revised 4 April2006 accepted 5 April 2006 doi Murine Atg8L Apg8L has significant homology with the other known mammalian Atg8 homologs LC3 GABARAP and GATE-16. However it is unclear whether murine Atg8L modification is mediated by human Atg4B Atg7 and Atg3. Expression of Atg8L in HEK293 cells led to cleavage of its C-terminus. In vitro the C-terminus of Atg8L was cleaved by human Atg4B but not human Atg4A or Atg4C. Atg8L-I formed an E1-substrate intermediate with Atg7C572S and an E2-substrate intermediate with Atg3C264S. A modified form of Atg8L was detected in the pelletable fraction in the presence of lysosomal protease inhibitors under nutrient-rich conditions. Cyan fluorescent protein CFP -Atg8L colocalized with yellow fluorescent protein YFP -LC3 in HeLa cells in the presence of the inhibitors. However little accumulation of the modified form of Atg8L was observed under conditions of starvation. These results indicate that Atg8L is the fourth modifier of mammalian Atg8 conjugation. Ubiquitylation and ubiquitylation-like reactions are post-translational modifications that play indispensable roles in many cellular events. Atg8 Apg8 Aut7 is a ubiquitin-like Ubl protein essential for autophagy in the yeast Saccharomyces cerevisiae 1 . Genetic analyses of yeast ATG gene mutants have suggested that the C-terminus of Atg8 is cleaved .

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