Isolation and characterization of AAP1. A gene encoding an alanine/arginine aminopeptidase in yeast.
The yeast AAP1 gene, encoding a putative amino-peptidase, 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.
Caprioglio
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Biology Department, University of New Mexico, Albuquerque 87131.
Padilla
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Werner-Washburne
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D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D018826:CD13 Antigens; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004271:DNA, Fungal; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D000626:Aminopeptidases; D018826:CD13 Antigens; C019721:aminopeptidase B