The yeast ISN1 (YOR155c) gene encodes a new type of IMP-specific 5'-nucleotidase.
BACKGROUND
The purine salvage enzyme inosine 5'-monophosphate (IMP)-specific 5'-nucleotidase catalyzes degradation of IMP to inosine. Although this enzymatic activity has been purified and characterized in Saccharomyces cerevisiae, the gene encoding IMP 5'-nucleotidase had not been identified.


RESULTS
Mass spectrometry analysis of several peptides of this enzyme purified from yeast allowed identification of the corresponding gene as YOR155c, an open reading frame of unknown function, renamed ISN1. The deduced Isn1p sequence was clearly not homologous to 5'-nucleotidases from other species. However, significant similarities to Isn1p were found in proteins of unknown function from Neurospora crassa, Plasmodium falciparum and several yeast species. Knock-out of ISN1 resulted in the total loss of IMP-specific 5'-nucleotidase activity, thus confirming that the ISN1 gene indeed encodes the enzymatic activity purified from yeast. In vivo studies revealed that, when IMP is overproduced through constitutive activation of the IMP de novo synthesis pathway, ISN1 is required for excretion of inosine and hypoxanthine in the medium.


CONCLUSIONS
We have identified a new yeast gene, ISN1 (YOR155c), as encoding IMP-specific 5'-nucleotidase activity. The ISN1 gene defines a new type of 5'-nucleotidase which was demonstrated to be functional in vivo.
Itoh
Roichi
R
Tokyo Kasei Gakuin University, 2600 Aiharamachi, Machida-city, Tokyo, Japan 194-0292. roytoh@kasei-gakuin.ac.jp
Saint-Marc
Christelle
C
Chaignepain
Stéphane
S
Katahira
Riko
R
Schmitter
Jean-Marie
JM
Daignan-Fornier
Bertrand
B
D015720:5'-Nucleotidase / Q000145:classification / Q000378:metabolism*; D000595:Amino Acid Sequence; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D029701:Saccharomyces cerevisiae Proteins; D010744:Phosphoric Monoester Hydrolases; D015720:5'-Nucleotidase