Isolation and characterization of AAP1. A gene encoding an alanine/arginine aminopeptidase in yeast

• Published in: The Journal of biological chemistry Vol. 268; no. 19; pp. 14310 - 14315
• Main Authors: Caprioglio, D.R., Padilla, C., Werner-Washburne, M.
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 05.07.1993
• Subjects: alanina; alanine; Amino Acid Sequence; amino acid sequences; aminopeptidases; Aminopeptidases - biosynthesis; Aminopeptidases - genetics; Animals; arginina; arginine; Base Sequence; beta-Galactosidase; beta-Galactosidase - biosynthesis; beta-Galactosidase - genetics; biosynthesis; carbohydrate metabolism; CD13 Antigens; chromosome; Chromosome Mapping; chromosomes; Chromosomes, Fungal; cromosomas; DNA, Fungal; DNA, Fungal - genetics; enzymology; gene; gene location; genes; Genes, Fungal; Genetic Complementation Test; genetic transformation; genetica; genetics; genetique; glicogeno; glycogen; glycogene; growth & development; Humans; localisation de gene; localizacion de genes; metabolism; metabolisme des glucides; metabolismo de carbohidratos; Mice; Molecular Sequence Data; mutant; mutantes; mutants; nucleotide sequence; nucleotide sequences; Open Reading Frames; peptidasas; peptidase; peptidases; Phenotype; Rats; Recombinant Fusion Proteins; Recombinant Fusion Proteins - biosynthesis; Restriction Mapping; RNA, Messenger; RNA, Messenger - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - growth & development; secuencia nucleica; Sequence Homology, Amino Acid; sequence nucleique; structural genes; transformacion genetica; transformation genetique
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)85242-0

The yeast AAP1 gene, encoding a putative aminopeptidase, was isolated based on its ability to suppress the temperature-sensitive growth on nonfermentable carbon sources of spr5, a stationary phase regulatory mutant. AAP1 was physically mapped to chromosome VIII between PUT2 and CUP1. Sequence analysis of the AAP1 gene showed a 1581-nucleotide open reading frame capable of encoding a 59-kilodalton protein. The protein encoded by this open reading frame exhibits approximately 40% sequence identity to human, rat, and mouse aminopeptidases. In limited regions, sequence identity between Aap1 and the mammalian aminopeptidases ranges from 53% to 93%. Insertional inactivation of the AAP1 gene resulted in a decrease in glycogen accumulation and the loss of the major band of arginine/alanine aminopeptidase activity. Strains carrying the AAP1 gene on a high copy plasmid show an increase in the major arginine/alanine aminopeptidase activity, a dramatic increase in glycogen accumulation, and an increase in transcription from a vector carrying lacZ fused to the promoter of a gene (SSA3) expressed during post-diauxic and stationary phases of the culture cycle. We conclude that although the AAP1 gene is not essential for viability, the AAP1 protein positively affects glycogen accumulation in yeast.