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Eukaryotic Cell, February 2006, p. 313-320, Vol. 5, No. 2
1535-9778/06/$08.00+0     doi:10.1128/EC.5.2.313-320.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Function and Properties of the Azf1 Transcriptional Regulator Change with Growth Conditions in Saccharomyces cerevisiae{dagger}

Matthew G. Slattery,1 Dritan Liko,2 and Warren Heideman1,2*

Pharmaceutical Sciences, School of Pharmacy,1 Department of Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin2

Received 29 September 2005/ Accepted 3 December 2005

Azf1 activates CLN3 transcription in Saccharomyces cerevisiae cells growing in glucose. Paradoxically, other studies have shown Azf1 to be almost undetectable in glucose-grown cells. Microarray experiments showed that Azf1 activates nonoverlapping gene sets in different carbon sources: in glucose, Azf1 activates carbon and energy metabolism genes, and in glycerol-lactate, Azf1 activates genes needed for cell wall maintenance. Consistent with the decreased expression of cell wall maintenance genes observed with azf1{Delta} mutants, we observed a marked growth defect in the azf1{Delta} cells at 37°C in nonfermentable medium. Cell wall integrity assays, such as sensitivity to calcofluor white, sodium dodecyl sulfate, or caffeine, confirmed cell wall defects in azf1{Delta} mutants in nonfermentable medium. Gel shift experiments show that Azf1 binds to DNA elements with the sequence AAAAGAAA (A4GA3), a motif enriched in the promoters of Azf1-sensitive genes and predicted by whole-genome studies. This suggests that many of the Azf1-dependent transcripts may be regulated directly by Azf1 binding. We found that the levels of Azf1 protein in glucose-grown cells were comparable to Azf1 levels in cells grown in glycerol-lactate; however, this could only be demonstrated with a cell extraction procedure that minimizes proteolysis. Glucose produces conditions that destabilize the Azf1 protein, a finding that may reflect a glucose-induced change in Azf1 tertiary or quaternary structure.

* Corresponding author. Mailing address: School of Pharmacy, University of Wisconsin, 425 N. Charter Street, Madison, WI 53706. Phone: (608) 262-1795. Fax: (608) 262-3397. E-mail: wheidema{at}facstaff.wisc.edu.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, February 2006, p. 313-320, Vol. 5, No. 2
1535-9778/06/$08.00+0     doi:10.1128/EC.5.2.313-320.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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