No abstract

The fructose-l,6-bisphosphatase [Fru(l ,6)P2ase] gene of the budding yeast, Kluyveromyces luctis, was cloned and sequenced. The gene encodes one open reading frame predicting a 354-amino-acid polypeptide. The polypeptide is different from other Fru(l,6)P2ases in that it contains a short amino-acid-insert region close to a basic residue located at the binding site for the allosteric inhibitor AMP. Comparison of the biochemical properties of the K. Zuctis enzyme with its closest homolog, the Succhuromyces cerevisiue Fru(1 ,6)P2ase (74% amino acid identity), reveals that the K. luctis enzyme is significantly less sensitive to AMP (Kl = 540 pM) than the S. cerevisiue enzyme (K,= 190 pM). However, studies with a K. luctis Fru(l,6)Pzase mutant, in which the insert region (amino acids 152-160) was deleted by site-directed mutagenesis [(des-152-160)Fru(l,6)Pzase], showed that the mutant enzyme had higher sensitivity to AMP inhibition (Kl = 280 pM) than the control K. luctis enzyme. Thus, the nine-amino-acid insert region appears to be responsible for the decreased AMP inhibition shown by the K. luctis wild-type enzyme. Catabolite-repression and catabolite-inactivation studies show that, unlike the complete repression of FBPI mRNA and inactivation of enzyme activity by glucose seen in S. cerevisiue, mRNA levels and enzyme activity of K. Zuctis Fru(16)P2ase decreased only about 2-4-fold due to the presence of glucose in the cell-culture medium.

Stereochemical Analysis of the Reaction Catalyzed by Yeast Protein Farnesyltransferase. --(EDELSTEIN, R. L.; WELLER, V. A.; DISTEFANO, M. D.; TUNG, J. S.; J.