Yeast AMP deaminase. Catalytic activity in Schizosaccharomyces pombe and chromosomal location in Saccharomyces cerevisiae.
The AMP deaminase gene was mapped to chromosome XIII of Saccharomyces cerevisiae strain JM1901. The AMP deaminase gene is located near SUP5, GAL80, SUF7, and SUF22. The presence of AMP deaminase in the fission yeast Schizosaccharomyces pombe was examined by comparing DNA hybridization, protein immunoreactivity, and catalytic activity from S. cerevisiae, known to contain the protein, to S. pombe. DNA hybridization experiments using the cloned S. cerevisiae AMP deaminase gene failed to hybridize to the genomic DNA from S. pombe strain 972h-s. Protein extracts from S. pombe and S. cerevisiae were analyzed in parallel and exhibited comparable AMP deaminase activities. Analysis of reaction intermediates in cell extracts of S. pombe established that IMP is formed directly from AMP without intervening steps. The AMP deaminase of S. pombe was purified 1,100-fold to a specific catalytic activity of 67 mumol/min/mg of protein. Purified protein interacted weakly with polyclonal antibodies prepared against S. cerevisiae AMP deaminase. AMP deaminases from both S. cerevisiae and S. pombe were activated by ATP with micromolar activation constants, are inhibited by coformycin, and are specific for AMP when compared to other purine nucleosides and nucleotides. The results establish that S. pombe contains an AMP deaminase with catalytic properties similar to that from S. cerevisiae, even though the DNA sequences of the genes and the immunoreactivity of the protein from S. pombe differs considerably from the AMP deaminase of S. cerevisiae. Genetic analysis of the pathways of purine metabolism in S. pombe (Pourquié, J., and Heslot, H. (1971) Genet. Res. 18, 33-44) had indicated the absence of AMP deaminase. The presence of a regulated AMP deaminase in S. pombe supports the hypothesis that eukaryotes regulate adenine nucleotide pools by the activity of AMP deaminase.
Sollitti
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P
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York 10461.
Merkler
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DJ
Estupiñán
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B
Schramm
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VL
D000659:AMP Deaminase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000378:metabolism; D015153:Blotting, Western; D002384:Catalysis; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D004271:DNA, Fungal; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D007291:Inosine Monophosphate / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D012568:Schizosaccharomyces / Q000201:enzymology*
D004271:DNA, Fungal; D007291:Inosine Monophosphate; D000249:Adenosine Monophosphate; D000659:AMP Deaminase