|Structural and functional analyses of APG5, a gene involved in autophagy in yeast|
Kametaka S, Matsuura A, Wada Y, Ohsumi Y
Category: autophagy ¤ Added: Feb 20, 2006 ¤ Rating: ◊◊
The APG5 gene of Saccharomyces cerevisiae was cloned from a yeast genomic library by complementation of autophagy defective phenotype of apg5-1 mutant. Structural analysis of the obtained genomic fragment showed that the APG5 gene encodes a novel hydrophilic protein of 294 amino-acid residues without apparent structural similarities to other proteins in the database. To examine its function, a null allele for APG5 (Aapg5) was constructed and introduced into yeast. Aapg5 cells germinated and grew normally in nutrient-rich condition, however, their viability reduced significantly upon the nutrient starvation. They were also shown to be defective in autophagy: they could not sequester autophagic bodies in the vacuole under nitrogen-starvation conditions. These phenotypes are identical to those found in the apg5-1 mutant. The lack of apparent phenotype in rich medium suggests that APG5 function is required only under nutrient starvation condition, however, Northern blot analysis showed that its expression levels remained unchanged after nutrient depletion.