ERG10 from Saccharomyces cerevisiae encodes acetoacetyl-CoA thiolase.
Two recessive alleles of ERG10 and three temperature-sensitive recessive alleles of HMG1 (3-hydroxy-3-methyl-glutaryl-CoA reductase isoenzyme 1) were isolated in a screen for mevalonate auxotrophs in Saccharomyces cerevisiae. The essential, single-copy ERG10 gene was cloned by complementation of the temperature-sensitive phenotype of erg10-21. The 1,194-base pair continuous open reading frame, encoding a 398-amino acid polypeptide with a calculated molecular mass of 41,681 daltons, was demonstrated to encode cytoplasmic aceto-acetyl-CoA thiolase. Acetoacetyl-CoA thiolase activity corresponded to the number of copies of ERG10 present in cell extracts, and null alleles of ERG10 produced no detectable acetoacetyl-CoA thiolase enzyme activity. The deduced amino acid sequence was 40-95% identical to acetoacetyl-CoA thiolases from other organisms. This identity included the active site cysteines located at amino acids 91 and 384 in the Erg10 protein.
Hiser
L
L
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Basson
M E
ME
Rine
J
J
D000101:Acetyl-CoA C-Acetyltransferase / Q000235:genetics*; D000483:Alleles; D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D016384:Consensus Sequence; D003593:Cytoplasm / Q000201:enzymology; D004271:DNA, Fungal / Q000235:genetics; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008798:Mevalonic Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D004271:DNA, Fungal; D000101:Acetyl-CoA C-Acetyltransferase; D008798:Mevalonic Acid