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Proteolytic activities in yeast after UV irradiation

I. Variation in proteinase levels in repair proficient Rad +strains

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Summary

Specific proteolytic activities are known to be induced in Escherichia coli following irradiation. Consequently it seemed of interest to investigate whether variations in proteinase activities occur in yeast.

Among the five most well known proteinases of Saccharomyces cerevisiae, we have found that proteinase B activity increases up to three times in wild-type RAD +yeast cells after a dose of 50 Jm-2 of 254 nm ultraviolet light (40% survival). Carboxypeptidase Y and aminopeptidase I (leucin aminopeptidase) activities were only moderately increased. Proteinase A activity was only slightly enhanced, while aminopeptidase II (lysin aminopeptidase) was unaffected in both RAD +strains studied.

The observed post UV-increase in proteinase B activity was inhibited by cycloheximide and was dose dependent. Increases in proteinase B levels were independent of the activation method used to destroy the proteinase B-inhibitor complex present in the crude yeast extracts.

A standard method for comparison of the postirradiation levels among different proteinases, strains and methods of activation is presented.

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Abbreviations

UV:

Ultraviolet

BRIJ-35:

Polyoxyethylene-23-lauryl ether

EDTA:

Ethylene diamine tetraacetic acid

EGTA:

Ethylene glycol bis (β-aminoethyl ether) tetraacetic acid

MOPS:

3-[N-morpholine]propansulfonic acid

HEPES:

N-2-Hydroxyethylpiperazine-N′-2-ethansulfonic acid

Tris:

Tris(hydroxy methyl)amino methane

BTPNA:

N-benzoyl-L-tyrosine-p-nitroanilide

CP.Y:

Carboxypeptidase Y

Leu.AP:

Leucin amino peptidase

Lys.AP:

Lysin amino peptidase

DMFA:

Dimethyl formamide

CHX:

Cycloheximide

PMSF:

Phenylmethyl sulfonyl fluoride

TCA:

Trichloroacetic acid

EC. 3.4.23.8:

Proteinase A

EC. 3.4.22.9:

Proteinase B

EC. 3.4.12.8:

Carboxypeptidase Y

EC. 3.4.24.4:

Thermolysin

EC. 3.4.23.1:

Pepsin A

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Authors and Affiliations

  1. Laboratoire d'Enzymologie du C.N.R.S., F-91190, Gif-sur-Yvette, France

    Jaime Schwencke

  2. Institut Curie, Section de Biologie, Centre Universitaire, Bât. 110, F-91405, Orsay, France

    Ethel Moustacchi

Authors
  1. Jaime Schwencke
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  2. Ethel Moustacchi
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Communicated by H. Böhme

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Schwencke, J., Moustacchi, E. Proteolytic activities in yeast after UV irradiation. Molec. Gen. Genet. 185, 290–295 (1982). https://doi.org/10.1007/BF00330800

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  • DOI: https://doi.org/10.1007/BF00330800

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