The proteinase yscB inhibitor (<i>PBI2</i>) gene of yeast and studies on the function of its protein product

Schu, Peter; Rendueles, Paz Suárez; Wolf, Dieter H.

doi:10.1111/j.1432-1033.1991.tb15874.x

Cited by 28 publications

(23 citation statements)

References 36 publications

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“…cubafion (Fig, :S). Under the same conditions the In= inhibitor mutant showed a constantly increased protein degradation rate of about 20-50% [12]. The mutant deficient in both inhibitors has the same protein desra.…”

Section: Protein Degradationmentioning

confidence: 64%

“…1). The amino acid sequence deduced from the nucleotide sequence is identical with the published primary structure of the protein, which had been determined by amino acid sequencing [12]. Ten of the 200 colonies of each transformation experiment were used for the test of the stability of the LEU2 F rototrophic phenotype.…”

Section: Isolation and Sequencing Of The Pal3 Genementioning

confidence: 86%

“…These spores were used to generate homozygous t~.i, ploids, deficient in the I^.a inhibitor and in both inhibitors, I^3 a~ttl I"2; Mating, sporulation and germination of the sogenerated strains and spores were not altered compared to the cells of the otherwise isogenic wild-type Strain (not shown). The activities of proteinase yscB and of the proteinase yscB inhibitor It)z increase under heat-stress conditions at 37°C [12,44]. The absence of the In2 inhibitor, however, does not affect the viability of the cells ur~der acute heat-stress conditions [12].…”

Section: Mata To Mata By the Use Of Ho Gene (See Section 2)mentioning

confidence: 99%

“…The activities of proteinase yscB and of the proteinase yscB inhibitor It)z increase under heat-stress conditions at 37°C [12,44]. The absence of the In2 inhibitor, however, does not affect the viability of the cells ur~der acute heat-stress conditions [12]. We tested the cell viability of the various inhibitor mutant strains under the combination of heat-stress anti starvation conditions.…”

Section: Mata To Mata By the Use Of Ho Gene (See Section 2)mentioning

confidence: 99%

“…The exact removal of the structural gene was demonstrated by sequencing, After insertion of the LEU2 gene [33] into the mutated DNA, yeast strains YSI8 [24] and YPS [12] were transformed with 5 l~g DNA of the linearized construct and the transformants vcere selected on MVplates without leucine, Clones were tested for inhibitory activity against purified proteinase A [15,34],…”

Section: 5 Isolation Sequertcing and Deletion Of The Pal 3 Genementioning

confidence: 99%

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The proteinase yscA‐inhibitor, I^A₃, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

Schu¹,

Wolf²

1991

FEBS Letters

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i'h¢ gen¢ of" the proteina~ y~cA inhibitor i":j, PAl3, of the yeast Saceharan)yce,~ ¢¢ecvi~ta¢ was isolated by oligonucl¢otidc screening of a genomic DNA library and sequenced. The gent codes for a single protein of 68 amino acids, The structural PAI3 gen¢ was deleted in vitro by oli$onucleotidesite.directed mutagenests. The mutated allele was introduced via homologous recombination into the 8enome of wild,type yeast and into the Itenome of a yeast mutant, which lacks the s~ond cytoplasmic proteina~-inhibitor, l°a, The deficiency of either or of both inhibitors has no effect on th© cell viability under various physiological conditions, The inhibitor mutants, ho~vever, show an increase in the lleneral in vivo protein degradation rate, The 1^3 mutant has a 2-~l.fold increased protein degradation rate in the tint 6 h aft©r a shift from rich medium onto starvation-medium, whereas the IBz mutant shows a constantly incrca~d degradation rate of 20~50~ under the same conditions. The inhibitor double null mutant has the same protein degradation rate as the [^~ null mutant. The~ results suggest an in vivo interaction b~tw©en the vacuolor endopeptidases and their cytoplasmic inhibitors.Pt~nt¢olysis; Prot¢ina,~ inhibitor; [^~; [no; Null mutant; Saccharomycts cerevisia¢ 1, INTRODUCTIONThe vacuole (lysosome) of the yeast is an actdic compartment, which contains multiple hydrolases (see reviews [1][2][3][4]). It is also a storage organelle for several compounds like phosphate, amino acids and calcium. The budding yeast Saccharomyces cerevisiae contains 1-2 large vacuoles per cell, which undergo multiple fu. sion and fragmentation processes during cell growth and sporulation (for review, see [5]).The two vacuolar endopeptidases proteinase yscA and yscB are essential enzymes for protein degradation during vegetative growth and, most pronounced, under starvation conditions ([6], for review see [1][2][3][4]). Proteinase yscA is even vital for cell viability under starvetion conditions [6]. The peptidases proteinase yscA, proteinase yscB and earboxypeptidase yscY are necessary for the differentiation prbcess of sporulation [6].The cytoplasm of the yeast S:tccharomyces cerevisiae contains the peptide inhibitors, Ia3, In2 and I c, specific for the vacuolar endopeptida.,+es proteinase yscA, proteinase yscB and carbo×ypeptidase yscY, respectively [7,8] No proteinase activities can be detected in cell extracts of wild-type yeast due to the formation of the enzyme/inhibitor complexes. The proteinase act ivities can be recovered by incubation at pH 5 or in the presence of detergents [13][14][15][16]. The inhibitors were found in the cytoplasmic fraction and only traces were found in the microsomal fraction [7]. The proteinase inhibitors are small and heat stable proteins (IA~: 7.6 kDa; In2:8.5 kDa), which contain no disulphide bonds [17][18][19][20]. Little is known about the function of the cytoplasmic proteinase inhibitors.The intracellular distribution of the proteinases and their inhibitors, as well as the observation that they are ex...

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Section: Protein Degradationmentioning

confidence: 64%

Section: Isolation and Sequencing Of The Pal3 Genementioning

confidence: 86%

Section: Mata To Mata By the Use Of Ho Gene (See Section 2)mentioning

confidence: 99%

Section: Mata To Mata By the Use Of Ho Gene (See Section 2)mentioning

confidence: 99%

Section: 5 Isolation Sequertcing and Deletion Of The Pal 3 Genementioning

confidence: 99%

See 3 more Smart Citations

The proteinase yscA‐inhibitor, I^A₃, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

Schu¹,

Wolf²

1991

FEBS Letters

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Proteinase Inhibitors

Laing

McManus

2018

Annual Plant Reviews Online

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Detection of Proteinases inSaccharomyces cerevisiaeby Flow Cytometry

Hutter

Miedl

Kuhmann

et al. 2005

Journal of the Institute of Brewing

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The physiological state of a yeast population used for inoculation determines how rapidly the cells adapt to new environmental conditions, begin proliferating and utilising extract. The decision as to whether a yeast culture is suitable for re-pitching should not be based only on viability determinations since this can be misleading. Increased proteolytic activity in a yeast population indicates the onset of senescence. A flow cytometric method has been developed for measuring a wide variety of proteinases in Saccharomyces cerevisiae employing a commercially available casein-dye conjugate. The detection of intracellular proteinase activity gives an early indication of apoptotic events and allows improved assessment of the physiological state of a yeast population. This knowledge will assist the industry to optimize the selection of yeast and its subsequent fermentation performance. Yeast cell autolysis with all its negative consequences for beer quality and stability will thus be minimised.

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The proteinase yscB inhibitor (PBI2) gene of yeast and studies on the function of its protein product

Cited by 28 publications

References 36 publications

The proteinase yscA‐inhibitor, I^A₃, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

The proteinase yscA‐inhibitor, I^A₃, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

Proteinase Inhibitors

Detection of Proteinases inSaccharomyces cerevisiaeby Flow Cytometry

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The proteinase yscB inhibitor (PBI2) gene of yeast and studies on the function of its protein product

Cited by 28 publications

References 36 publications

The proteinase yscA‐inhibitor, IA3, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

The proteinase yscA‐inhibitor, IA3, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

Proteinase Inhibitors

Detection of Proteinases inSaccharomyces cerevisiaeby Flow Cytometry

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The proteinase yscA‐inhibitor, I^A₃, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology

The proteinase yscA‐inhibitor, I^A₃, gene Studies of cytoplasmic proteinase inhibitor deficiency on yeast physiology