Identification of direct and indirect targets of the Gln3 and Gat1 activators by transcriptional profiling in response to nitrogen availability in the short and long term.
Nitrogen catabolite repression (NCR) consists in the specific inhibition of transcriptional activation of genes encoding the permeases and catabolic enzymes needed to degrade poor nitrogen sources. Under nitrogen limitation or rapamycin treatment, NCR genes are activated by Gln3 or Gat1, or by both factors. To compare the sets of genes responding to rapamycin or to nitrogen limitation, we used DNA microarrays to establishing the expression profiles of a wild type strain, and of a double gln3Delta-gat1Delta strain, grown on glutamine, after addition of rapamycin, on proline, or after a shift from glutamine to proline. Analysis of microarray data revealed 392 genes whose expression was dependent on the nitrogen source quality. 91 genes were activated in a GATA factor-dependent manner in all growth conditions, suggesting a direct role of Gln3 and Gat1 in their expression. Other genes were only transiently up-regulated (stress-responsive genes) or down-regulated (genes encoding ribosomal proteins and translational factors) upon nitrogen limitation, and this regulation was delayed in a gln3Delta-gat1Delta strain. Repression of amino acid and nucleotide biosynthetic genes after a nitrogen shift did not depend on Gcn4. Several transporter genes were repressed as a consequence of enhanced levels of NCR-responsive permeases present at the plasma membrane.
Scherens
Bart
B
Institut de Recherches Microbiologiques J-M Wiame, and Laboratoire de Microbiologie, Université Libre de Bruxelles, Brussels, Belgium.
Feller
André
A
Vierendeels
Fabienne
F
Messenguy
Francine
F
Dubois
Evelyne
E
D001665:Binding Sites; D050980:GATA Transcription Factors / Q000378:metabolism / Q000502:physiology*; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D005973:Glutamine / Q000378:metabolism; D005979:Glutathione Peroxidase; D009584:Nitrogen / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D011328:Prions / Q000502:physiology; D011392:Proline / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000502:physiology*; D012269:Ribosomal Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D020123:Sirolimus / Q000494:pharmacology; D014157:Transcription Factors / Q000502:physiology*
C096100:GAT1 protein, S cerevisiae; D050980:GATA Transcription Factors; C071664:GLN3 protein, S cerevisiae; D011328:Prions; D012097:Repressor Proteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D005973:Glutamine; D011392:Proline; D005979:Glutathione Peroxidase; C067020:URE2 protein, S cerevisiae; D009584:Nitrogen; D020123:Sirolimus