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J Biol Chem, Vol. 273, Issue 21, 12696-12702, May 22, 1998
From the Institut für Mikrobiologie und Genetik, Abteilung
für Molekulare Mikrobiologie, Georg-August-Universität,
Grisebachstrasse 8, D-37077 Göttingen, Germany
In Saccharomyces cerevisiae the
GCN4 gene encodes the transcriptional activator of the
"general control" system of amino acid bioynthesis, a network of at
least 12 different biosynthetic pathways. We characterized the
consequences of the general control response upon the signal "amino
acid starvation" induced by the histidine analogue 3-aminotriazole
with respect to Gcn4p levels in more detail. Therefore, we established
test systems to monitor the time course of different parameters,
including GCN4 mRNA, Gcn4 protein, Gcn4p DNA binding
activity, as well as Gcn4p transactivation ability. We observed a
biphasic response of Gcn4p activity in the cell. At first, translation
of GCN4 mRNA is induced within 20 min after switch to
starvation conditions. However, an additional increase in
GCN4 transcript steady state level was observed, leading to
an additional second phase of GCN4 expression after 3-4 h
of starvation. The DNA binding activity of Gcn4p, as well as the ability to activate transcription of target genes, correlate with the
amount of Gcn4 protein in the cell, suggesting that under the tested
conditions there is no additional regulation of DNA binding or
transactivation ability of Gcn4p, respectively.
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