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Conformational changes play a role in regulating the activity of the proline utilization pathway‐specific regulator in Saccharomyces cerevisiae
Author(s) -
Des Etages Shelley Ann G.,
Saxena Deepti,
Huang Hoching L.,
Falvey Darlene A.,
Barber Dianna,
Brandriss Marjorie C.
Publication year - 2001
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.2001.02432.x
Subject(s) - proline , saccharomyces cerevisiae , biology , regulator , biochemistry , amino acid , protease , yeast , inducer , mutation , gene , conformational change , microbiology and biotechnology , enzyme
In Saccharomyces cerevisiae , the ability to use proline as a nitrogen source requires the Put3p transcriptional regulator, which turns on the expression of the proline utilization genes, PUT1 and PUT2 , in the presence of the inducer proline and in the absence of preferred nitrogen sources. Changes in target gene expression occur through an alteration in activity of the DNA‐bound Put3p, a member of the Zn(II) 2 Cys 6 binuclear cluster family of proteins. Here, we report that the ‘on’ conformation can be mimicked in the absence of proline by the insertion of an epitope tag in several different places in the protein, as well as by specific amino acid changes that suppress a put3 mutation leading to non‐inducibility of the pathway. In addition, the presence of proline causes a conformational change in the Put3 protein detected by increased sensitivity to thrombin or V8 protease. These findings suggest that Put3p shifts from an inactive to an activate state via conformational changes.