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Inhibition of ubiquitin/proteasome‐dependent proteolysis in Saccharomyces cerevisiae by a Gly‐Ala repeat
Author(s) -
Heessen Stijn,
Dantuma Nico P.,
Tessarz Peter,
Jellne Marianne,
Masucci Maria G.
Publication year - 2003
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(03)01296-1
Subject(s) - saccharomyces cerevisiae , proteolysis , yeast , ubiquitin , proteasome , biology , biochemistry , recombinant dna , ubiquitin ligase , fusion protein , protein degradation , microbiology and biotechnology , chemistry , gene , enzyme
The glycine‐alanine (GA) repeat of the Epstein–Barr virus nuclear antigen‐1 inhibits in cis ubiquitin‐dependent proteolysis in mammalian cells through a yet unknown mechanism. In the present study we demonstrate that the GA repeat targets an evolutionarily conserved step in proteolysis since it can prevent the degradation of proteasomal substrates in the yeast Saccharomyces cerevisiae . Insertion of yeast codon‐optimised recombinant GA (rGA) repeats of different length in green fluorescent protein reporters harbouring N‐end rule or ubiquitin fusion degradation signals resulted in efficient stabilisation of these substrates. Protection was also achieved in rpn10Δ yeast suggesting that this polyubiquitin binding protein is not required for the rGA effect. The conserved effect of the GA repeat in yeast opens the possibility for the use of genetic screens to unravel its mode of action.