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A new protein conjugation system in human. The counterpart of the yeast Apg12p conjugation system essential for autophagy.

Autophagy is an intracellular process for bulk degradation of cytoplasmic components. We recently found a protein conjugation system essential for autophagy in the yeast, Saccharomyces cerevisiae. The C-terminal glycine of a novel modifier protein, Apg12p, is conjugated to a lysine residue of Apg5p via an isopeptide bond. This conjugation reaction is mediated by Apg7p, a ubiquitin activating enzyme (E1)-like enzyme, and Apg10p, suggesting that it is a ubiquitination-like system (Mizushima, N., Noda, T., Yoshimori, T., Tanaka, Y., Ishii, T., George, M. D., Klionsky, D. J., Ohsumi, M. , and Ohsumi, Y. (1998) Nature 395, 395-398). Although autophagy is a ubiquitous process in eukaryotic cells, no molecule involved in autophagy has yet been identified in higher eukaryotes. We reasoned that this conjugation system could be conserved. Here we report cloning and characterization of the human homologue of Apg12 (hApg12). It is a 140-amino acid protein and possesses 27% identity and 48% similarity with the yeast Apg12p, but no apparent homology to ubiquitin. Northern blot analysis showed that its expression was ubiquitous in human tissues. We found that it was covalently attached to another protein. This target protein was identified to be the human Apg5 homologue (hApg5). Mutagenic analyses suggested that this conjugation was formed via an isopeptide bond between the C-terminal glycine of hApg12 and Lys-130 of hApg5. These findings indicate that the Apg12 system is well conserved and may function in autophagy also in human cells.

Pubmed ID: 9852036 RIS Download

Mesh terms: Adult | Amino Acid Sequence | Animals | Autophagy | Autophagy-Related Protein 12 | Autophagy-Related Protein 5 | COS Cells | Cloning, Molecular | Fungal Proteins | Glycine | Humans | Lysine | Mice | Molecular Sequence Data | Organ Specificity | Proteins | RNA, Messenger | Recombinant Proteins | Saccharomyces cerevisiae | Saccharomyces cerevisiae Proteins | Sequence Alignment | Sequence Homology, Amino Acid | Transcription, Genetic | Transfection | Ubiquitin-Protein Ligases

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