A single point mutation results in a constitutively activated and feedback-resistant chorismate mutase of Saccharomyces cerevisiae.
The Saccharomyces cerevisiae ARO7 gene product chorismate mutase, a single-branch-point enzyme in the aromatic amino acid biosynthetic pathway, is activated by tryptophan and subject to feedback inhibition by tyrosine. The ARO7 gene was cloned on a 2.05-kilobase EcoRI fragment. Northern (RNA) analysis revealed a 0.95-kilobase poly(A)+ RNA, and DNA sequencing determined a 771-base-pair open reading frame capable of encoding a protein 256 amino acids. In addition, three mutant alleles of ARO7 were cloned and sequenced. These encoded chorismate mutases which were unresponsive to tyrosine and tryptophan and were locked in the on state, exhibiting a 10-fold-increased basal enzyme activity. A single base pair exchange resulting in a threonine-to-isoleucine amino acid substitution in the C-terminal part of the chorismate mutase was found in all mutant strains. In contrast to other enzymes in this pathway, no significant homology between the monofunctional yeast chorismate mutase and the corresponding domains of the two bifunctional Escherichia coli enzymes was found.
Schmidheini
T
T
Institute of Microbiology, Swiss Federal Institute of Technology, Zurich.
Sperisen
P
P
Paravicini
G
G
Hütter
R
R
Braus
G
G
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D002826:Chorismate Mutase / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D005246:Feedback; D005796:Genes; D005800:Genes, Fungal; D005838:Genotype; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D014443:Tyrosine / Q000494:pharmacology
D014443:Tyrosine; D007535:Isomerases; D002826:Chorismate Mutase