Polymorphism of the MPR1 gene required for toxic proline analogue resistance in the Saccharomyces cerevisiae complex species.
We recently discovered, on the chromosome of Saccharomyces cerevisiae sigma 1278b, novel MPR1 and MPR2 genes required for resistance to a toxic analogue of L-proline, L-azetidine-2-carboxylic acid. The MPR genes, which were absent in the S. cerevisiae genome project strain S288C, encoded a novel acetyltransferase of 229 amino acids that detoxifies the analogue by acetylating it. The MPR1 gene homologue found in Schizosaccharomyces pombe was also shown to encode a similar acetyltransferase. To further analyse the origin and the physiological role of the yeast novel gene, we report here the comparative analysis of the MPR1 gene in the S. cerevisiae complex spp. which belong to the Saccharomyces sensu stricto group. Only the type strain of S. paradoxus exhibited resistance and acetyltransferase activity to L-azetidine-2-carboxylic acid. PCR was then used to isolate the new MPR1 homologue (Spa MPR1) from S. paradoxus with the primers based on the sequence of the MPR1 gene. Gene expression and enzymatic analysis showed that the cloned Spa MPR1 gene encodes an L-azetidine-2-carboxylic acid acetyltransferase of 231 amino acids, which has 87% identity to the MPR1 protein. We also found in the protein databases that S. bayanus contains a DNA fragment that is partly homologous to the MPR1 gene. However, the gene product was considered to lose the enzymatic activity, possibly due to the gene truncation or the base substitution(s) at the important region for catalysis. Further, genomic PCR analysis showed that most of the S. cerevisiae complex spp. have the sequence highly homologous to the MPR1 gene.
Kimura
Yasuko
Y
Department of Bioscience, Fukui Prefectural University, Fukui 910-1195, Japan.
Nakamori
Shigeru
S
Takagi
Hiroshi
H
D000107:Acetylation; D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001383:Azetidinecarboxylic Acid / Q000494:pharmacology*; D001483:Base Sequence; D018797:Cell Cycle Proteins; D025141:Drug Resistance, Fungal; D005656:Fungal Proteins / Q000235:genetics*; D017353:Gene Deletion; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D011494:Protein Kinases; D012440:Saccharomyces / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins; D016415:Sequence Alignment; D013045:Species Specificity
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D029702:Schizosaccharomyces pombe Proteins; D001383:Azetidinecarboxylic Acid; D000123:Acetyltransferases; D011494:Protein Kinases; C413334:Mpr1 protein, S pombe