title: Cloning of two related genes encoding the 56-kDa and 123-kDa subunits of trehalose synthase from the yeast Saccharomyces cerevisiae.
abstract: Preparations of intact trehalose synthase contain three polypeptides with molecular masses of 56, 102 and 123 kDa. We have cloned the genes TSS1 and TSL1 coding for the 56- and 123-kDa subunits, respectively. These genes are located on chromosomes II (TSS1) and XIII (TSL1). The TSS1 gene was found to be identical with CIF1, a gene required for normal growth on glucose. The product of the entire TSS1 gene exhibits 37% identity with a 502-amino-acid stretch from the middle of the TSL1 product. Disruption of the TSS1 gene in yeast eliminates both trehalose 6-phosphate synthase (Tre6P synthase) and trehalose 6-phosphate phosphatase (Tre6Pase) activities, and reintroduction of this gene restores these activities. Transformation of Escherichia coli with TSS1 increases its Tre6P synthase activity. Specific proteolytic degradation of the 123-kDa polypeptide from the N-terminus greatly influences the Tre6P synthase activity, decreasing its inhibition by phosphate and activatability by fructose 6-phosphate but has little effect on the Tre6Pase activity. These results suggest that this N-terminal part confers regulatory properties upon the Tre6P synthase activity.
authors.affiliation: Research Laboratory, Alko Ltd, Helsinki, Finland.
authors.forename: O E
authors.initials: OE
authors.lastname: Vuorio
authors.forename: N
authors.initials: N
authors.lastname: Kalkkinen
authors.forename: J
authors.initials: J
authors.lastname: Londesborough
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal; D004789:Enzyme Activation; D004926:Escherichia coli; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D014357:Trypsin
chemical_list: D004271:DNA, Fungal; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase; C023178:trehalose-phosphatase; D010744:Phosphoric Monoester Hydrolases; D014357:Trypsin; D005947:Glucose