A high-affinity uptake system for branched-chain amino acids in Saccharomyces cerevisiae.
In order to isolate mutants with impaired uptake of branched-chain amino acids, mutants were induced which on complex medium were sensitive to an inhibitor of branched-chain amino acid biosynthesis. Eighteen mutants of independent origin were found. Ten of them were assayed for branched-chain amino acid uptake. Three of these were impaired in the uptake of L-valine, L-isoleucine and L-leucine, while the rest were unaffected in uptake of any of the three amino acids. Kinetics of the uptake by one selected mutant and the parental strain S288C were compared to models for one or two systems obeying Michaelis-Menten kinetics. This analysis suggested that a high-affinity system for all three amino acids is absent in the mutant, whereas low-affinity uptake of L-isoleucine and L-leucine by one or more systems remains unaffected. Moreover, medium-affinity uptake components for L-valine and L-leucine, not originally seen in the wild type, were identified in the mutant. In the wild type, 10 mM of any of the amino acids L-alanine, L-cysteine, L-isoleucine, L-leucine, L-tryptophan and L-valine reduce uptake of any of the three branched-chain amino acids. We propose that a permease responsible for high-affinity uptake of the branched-chain amino acids in strain S288C is partially or completely inactive in the mutant. Tetrad analysis shows that the phenotype can be ascribed to a single Mendelian gene. The wild-type allele is denoted BAP1 for branched-chain amino acid permease. The BAP1-dependent system is different from the general amino acid permease.
Tullin
S
S
Department of Yeast Genetics, Carlsberg Laboratory, Copenhagen Valby, Denmark.
Gjermansen
C
C
Kielland-Brandt
M C
MC
D000597:Amino Acids, Branched-Chain / Q000378:metabolism*; D001692:Biological Transport; D003470:Culture Media; D007532:Isoleucine / Q000378:metabolism; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014633:Valine / Q000378:metabolism
D000597:Amino Acids, Branched-Chain; D003470:Culture Media; D007532:Isoleucine; D007930:Leucine; D014633:Valine