title: The product of the his4 gene cluster in Saccharomyces cerevisiae. A trifunctional polypeptide.
abstract: The his4 region of yeast encodes the information for the third (phosphoribosyl-AMP cyclohydrolase), second (phosphoribosyl-ATP pyrophosphohydrolase), and tenth (histidinol dehydrogenase) steps in the histidine biosynthetic pathway. These three activities co-purify with a single protein which has a subunit molecular weight of 95,000 (95,000 protein), as determined by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate. Extracts of yeast strains which carry nonsense or deletion mutations in various portions of the his4 region, purified in parallel by affinity chromatography on AMP-agarose columns, were examined on sodium dodecyl sulfate-polyacrylamide gel electrophoresis slabs. All such mutant extracts examined were found to lack the 95,000 protein found in a strain carrying a wild type his4 allele. The presence of a protease inhibitor, phenylmethylsulfonyl fluoride, during the purification of the trifunctional enzyme prevented the degradation of the 95,000 protein to polypeptides of lower molecular weight. Monospecific antibody prepared against the 95,000 protein removed all three of the activities specified by his4 from solution; active 95,000 protein was recovered in the resuspended immunoprecipitates. All this evidence shows that the product of the his4 region is a trifunctional, 95,000-dalton protein. Preliminary evidence from two-dimensional gel electrophoresis, NH2-terminal analysis, and gel filtration column chromatography indicates that the native trifunctional enzyme is a dimer of identical 95,000-dalton subunits.
mesh_terms: D000249:Adenosine Monophosphate / Q000031:analogs & derivatives; D000429:Alcohol Oxidoreductases / Q000096:biosynthesis; D000619:Aminohydrolases / Q000096:biosynthesis; D005796:Genes; D006641:Histidinol; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000096:biosynthesis*; D009154:Mutation; D012267:Ribosemonophosphates; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
chemical_list: D009097:Multienzyme Complexes; D012267:Ribosemonophosphates; D000249:Adenosine Monophosphate; D006641:Histidinol; D000429:Alcohol Oxidoreductases; C021358:histidinol dehydrogenase; D000619:Aminohydrolases; C022383:phosphoribosyl-AMP cyclohydrolase