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The function and properties of the Azf1 transcriptional regulator change with growth conditions in Saccharomyces cerevisiae.
Eukaryot Cell. 2006 Feb; 5(2):313-20.EC

Abstract

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 azf1Delta mutants, we observed a marked growth defect in the azf1Delta cells at 37 degrees C in nonfermentable medium. Cell wall integrity assays, such as sensitivity to calcofluor white, sodium dodecyl sulfate, or caffeine, confirmed cell wall defects in azf1Delta 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.

Authors+Show Affiliations

Pharmaceutical Sciences, School of Pharmacy, University of Wisconsin, 425 N. Charter Street, Madison, WI 53706, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

16467472

Citation

Slattery, Matthew G., et al. "The Function and Properties of the Azf1 Transcriptional Regulator Change With Growth Conditions in Saccharomyces Cerevisiae." Eukaryotic Cell, vol. 5, no. 2, 2006, pp. 313-20.
Slattery MG, Liko D, Heideman W. The function and properties of the Azf1 transcriptional regulator change with growth conditions in Saccharomyces cerevisiae. Eukaryot Cell. 2006;5(2):313-20.
Slattery, M. G., Liko, D., & Heideman, W. (2006). The function and properties of the Azf1 transcriptional regulator change with growth conditions in Saccharomyces cerevisiae. Eukaryotic Cell, 5(2), 313-20.
Slattery MG, Liko D, Heideman W. The Function and Properties of the Azf1 Transcriptional Regulator Change With Growth Conditions in Saccharomyces Cerevisiae. Eukaryot Cell. 2006;5(2):313-20. PubMed PMID: 16467472.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The function and properties of the Azf1 transcriptional regulator change with growth conditions in Saccharomyces cerevisiae. AU - Slattery,Matthew G, AU - Liko,Dritan, AU - Heideman,Warren, PY - 2006/2/10/pubmed PY - 2006/5/9/medline PY - 2006/2/10/entrez SP - 313 EP - 20 JF - Eukaryotic cell JO - Eukaryot Cell VL - 5 IS - 2 N2 - 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 azf1Delta mutants, we observed a marked growth defect in the azf1Delta cells at 37 degrees C in nonfermentable medium. Cell wall integrity assays, such as sensitivity to calcofluor white, sodium dodecyl sulfate, or caffeine, confirmed cell wall defects in azf1Delta 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. SN - 1535-9778 UR - https://www.unboundmedicine.com/medline/citation/16467472/The_function_and_properties_of_the_Azf1_transcriptional_regulator_change_with_growth_conditions_in_Saccharomyces_cerevisiae_ DB - PRIME DP - Unbound Medicine ER -