PMID:12890687

Citation	Nemoto, T, Tanida, I, Tanida-Miyake, E, Minematsu-Ikeguchi, N, Yokota, M, Ohsumi, M, Ueno, T and Kominami, E (2003) The mouse APG10 homologue, an E2-like enzyme for Apg12p conjugation, facilitates MAP-LC3 modification. J. Biol. Chem. 278:39517-26
Abstract	Autophagy is a process for the bulk degradation of cytosolic compartments by lysosomes/vacuoles. The formation of autophagosomes involves a dynamic rearrangement of the membrane for which two ubiquitin-like modifications (the conjugation of Apg12p and the modification of a soluble form of MAP-LC3 to a membrane-bound form) are essential. In yeast, Apg10p is an E2-like enzyme essential for Apg12p conjugation. The isolated mouse APG10 gene product interacts with mammalian Apg12p dependent on mammalian Apg7p (E1-like enzyme), and facilitates Apg12p conjugation. The interaction of Apg10p with Apg12p is dependent on the carboxyl-terminal glycine of Apg12p. Mutational analysis of the predicted active site cysteine (Cys161) within mouse Apg10p shows that mutant Apg10pC161S, which can form a stable intermediate with Apg12p, inhibits Apg12p conjugation even in the presence of Apg7p, while overexpression of Apg7p facilitates formation of an Apg12p-Apg5p conjugate. Furthermore, the coexpression of Apg10p with Apg7p facilitates the modification of a soluble form of MAP-LC3 to a membrane-bound form, a second modification essential for autophagy. Mouse Apg10p interacts with MAP-LC3 in HEK293 cells, while no mutant Apg10pC161S forms any intermediate with MAP-LC3. Direct interaction between Apg10p and MAP-LC3 is also demonstrated by yeast two-hybrid analysis. The inability of mutant Apg10pC161S to form any intermediate with MAP-LC3 has ruled out the possibility that MAP-LC3 interacts with Apg10p as a substrate.
Links	PubMed Online version:10.1074/jbc.C200334200
Keywords	Amino Acid Sequence; Animals; Autophagy; Base Sequence; Cell Line; DNA, Complementary/genetics; DNA, Complementary/isolation & purification; Humans; Ligases/genetics; Ligases/metabolism; Mice; Microtubule-Associated Proteins/genetics; Microtubule-Associated Proteins/metabolism; Models, Biological; Molecular Sequence Data; Mutagenesis, Site-Directed; Oxidoreductases/genetics; Oxidoreductases/metabolism; Proteins/genetics; Proteins/metabolism; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Saccharomyces cerevisiae Proteins; Sequence Homology, Amino Acid; Two-Hybrid System Techniques; Ubiquitin-Conjugating Enzymes
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