Saccharomyces cerevisiae sigma 1278b has novel genes of the N-acetyltransferase gene superfamily required for L-proline analogue resistance.
We discovered on the chromosome of Saccharomyces cerevisiae Sigma 1278b novel genes involved in L-proline analogue L-azetidine-2-carboxylic acid resistance which are not present in the standard laboratory strains. The 5.4 kb-DNA fragment was cloned from the genomic library of the L-azetidine-2-carboxylic acid-resistant mutant derived from a cross between S. cerevisiae strains S288C and Sigma 1278b. The nucleotide sequence of a 4.5-kb segment exhibited no identity with the sequence in the genome project involving strain S288C. Deletion analysis indicated that one open reading frame encoding a predicted protein of 229 amino acids is indispensable for L-azetidine-2-carboxylic acid resistance. The protein sequence was found to be a member of the N-acetyltransferase superfamily. Genomic Southern analysis and gene disruption showed that two copies of the novel gene with one amino acid change at position 85 required for L-azetidine-2-carboxylic acid resistance were present on chromosomes X and XIV of Sigma 1278b background strains. When this novel MPR1 or MPR2 gene (sigma 1278b gene for L-proline analogue resistance) was introduced into the other S. cerevisiae strains, all of the recombinants were resistant to L-azetidine-2-carboxylic acid, indicating that both MPR1 and MPR2 are expressed and have a global function in S. cerevisiae.
Takagi
H
H
Department of Bioscience, Fukui Prefectural University, Matsuoka-cho, Fukui 910-1195, Japan. hiro@fpu.ac.jp
Shichiri
M
M
Takemura
M
M
Mohri
M
M
Nakamori
S
S
D000595:Amino Acid Sequence; D001191:Arylamine N-Acetyltransferase / Q000235:genetics*; D001383:Azetidinecarboxylic Acid / Q000494:pharmacology*; D001483:Base Sequence; D015139:Blotting, Southern; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular; D004352:Drug Resistance, Microbial / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D011392:Proline / Q000031:analogs & derivatives*; D011494:Protein Kinases; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins; D016415:Sequence Alignment
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D029702:Schizosaccharomyces pombe Proteins; D001383:Azetidinecarboxylic Acid; D011392:Proline; D001191:Arylamine N-Acetyltransferase; D011494:Protein Kinases; C413334:Mpr1 protein, S pombe