Molecular cloning and sequencing of genomic DNA encoding yeast vacuolar carboxypeptidase yscS.
A Saccharomyces cerevisiae genomic DNA encoding vacuolar carboxypeptidase yscS was cloned from a yeast YEp13 library by complementation of the previously characterized mutation cps1-1 [(1981) J. Bacteriol. 147, 418-426], by means of staining carboxypeptidase activity in yeast colonies. The nucleotide sequence of the cloned gene was determined. The open reading frame of CPS1 consists of 576 codons and therefore encodes a protein of 64961 molecular weight. A stretch of 19 residues near the N-terminus of the deduced polypeptide sequence contains characteristics common to known hydrophobic leader sequences. CPS1 was determined by DNA blot analysis to be a single copy gene located on chromosome X. The cloned fragment was used to identify a 2.1 kb mRNA. A transcriptional activation of CPS1 occurs when cells grow on a substrate of carboxy-peptidase yscS as sole nitrogen source.
Bordallo
J
J
Departamento de Biología Functional, Universidad de Oviedo, Spain.
Bordallo
C
C
Gascón
S
S
Suárez-Rendueles
P
P
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002268:Carboxypeptidases / Q000235:genetics*; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D012313:RNA / Q000032:analysis; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D004271:DNA, Fungal; D012313:RNA; D002268:Carboxypeptidases