The proteinase yscB inhibitor (PB12) gene of yeast and studies on the function of its protein product.
The gene for proteinase yscB inhibitor I2B (PBI2) from Saccharomyces cerevisiae was isolated by oligonucleotide screening of a genomic DNA library, and was sequenced. The gene codes for a single protein of 75 amino acids. In contrast to the published amino acid sequence [Maier, K., Müller, H., Tesch, R., Trolp, T., Witt, I. & Holzer, H. (1979) J. Biol. Chem. 254, 12,555-12,561] the DNA sequence revealed a valine instead of a leucine at position 33 (32 of the mature protein). Therefore the primary sequences of the isoinhibitors I2B of S. cerevisiae and I1B of Saccharomyces carlsbergensis differ only at position 34 (glutamic acid/lysine). The open reading frame of PBI2 was replaced in vitro by the URA3 gene and a I2B null mutant of S. cerevisiae was constructed by gene replacement. The mutation resulted in an elevation of the protein degradation rate by 50% when grown under nutritional stress compared to the isogenic wild type. Growth and viability of the cells was not significantly affected by the absence of I2B.
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Institut für Biochemie der Universität Stuttgart, Federal Republic of Germany.
Suarez Rendueles
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Wolf
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D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005800:Genes, Fungal; D005838:Genotype; D007700:Kinetics; D008969:Molecular Sequence Data; D012097:Repressor Proteins; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D012697:Serine Endopeptidases; D015842:Serine Proteinase Inhibitors / Q000378:metabolism* / Q000494:pharmacology; D033921:Vesicular Transport Proteins
D004271:DNA, Fungal; C068476:PBI2 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D015842:Serine Proteinase Inhibitors; D033921:Vesicular Transport Proteins; D012697:Serine Endopeptidases; C023302:yeast proteinase B