Role of the complex upstream region of the GDH2 gene in nitrogen regulation of the NAD-linked glutamate dehydrogenase in Saccharomyces cerevisiae.
We analyzed the upstream region of the GDH2 gene, which encodes the NAD-linked glutamate dehydrogenase in Saccharomyces cerevisiae, for elements important for the regulation of the gene by the nitrogen source. The levels of this enzyme are high in cells grown with glutamate as the sole source of nitrogen and low in cells grown with glutamine or ammonium. We found that this regulation occurs at the level of transcription and that a total of six sites are required to cause a CYC1-lacZ fusion to the GDH2 gene to be regulated in the same manner as the NAD-linked glutamate dehydrogenase. Two sites behaved as upstream activation sites (UASs). The remaining four sites were found to block the effects of the two UASs in such a way that the GDH2-CYC1-lacZ fusion was not expressed unless the cells containing it were grown under conditions favorable for the activity of both UASs. This complex regulatory system appears to account for the fact that GDH2 expression is exquisitely sensitive to glutamine, whereas the expression of GLN1, coding for glutamine synthetase, is not nearly as sensitive.
Miller
S M
SM
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Magasanik
B
B
D000595:Amino Acid Sequence; D000641:Ammonia / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal; D015966:Gene Expression Regulation, Fungal; D005969:Glutamate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism*; D014176:Protein Biosynthesis; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic
D004271:DNA, Fungal; D005971:Glutamates; D012331:RNA, Fungal; D012333:RNA, Messenger; D018698:Glutamic Acid; D000641:Ammonia; D005969:Glutamate Dehydrogenase; D009584:Nitrogen