Open AccessSaccharomyces cerevisiae Σ1278b Has Novel Genes of the N -Acetyltransferase Gene Superfamily Required for l -Proline Analogue Resistance
Author(s) -
Hiroshi Takagi,
Mika Shichiri,
Masashi Takemura,
Miho Mohri,
Shigeru Nakamori
Publication year - 2000
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.182.15.4249-4256.2000
Subject(s) - biology , saccharomyces cerevisiae , open reading frame , gene , genetics , genomic dna , nucleic acid sequence , mutant , microbiology and biotechnology , peptide sequence , genomic library , amino acid , biochemistry
We discovered on the chromosome ofSaccharomyces cerevisiae Σ1278b novel genes involved inl -proline analoguel -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 thel -azetidine-2-carboxylic acid-resistant mutant derived from a cross betweenS. cerevisiae strains S288C and Σ1278b. 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 forl -azetidine-2-carboxylic acid resistance. The protein sequence was found to be a member of theN -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 forl -azetidine-2-carboxylic acid resistance were present on chromosomes X and XIV of Σ1278b background strains. When this novelMPR1 orMPR2 gene (sigma 1278b gene forl -proline analogue resistance) was introduced into the otherS. cerevisiae strains, all of the recombinants were resistant tol -azetidine-2-carboxylic acid, indicating that bothMPR1 andMPR2 are expressed and have a global function inS. cerevisiae .