Expression of the yeast glycogen phosphorylase gene is regulated by stress-response elements and by the HOG MAP kinase pathway.
Yeast glycogen metabolism responds to environmental stressors such as nutrient limitation and heat shock. This response is mediated, in part, by the regulation of the glycogen metabolic genes. Environmental stressors induce a number of glycogen metabolic genes, including GPH1, which encodes glycogen phosphorylase. Primer extension analysis detected two start sites for GPH1, one of which predominated. Sequences upstream of these sites included a possible TATA element. Mutation of this sequence reduced GPH1 expression by a factor of 10 but did not affect start site selection. This mutation also did not affect the relative induction of GPH1 upon entry into stationary phase. Three candidates for stress response elements (STREs) were found upstream of the TATA sequence. Mutation of the STREs showed that they were required for regulation of GPH1 expression in early stationary phase, and in response to osmotic shock and heat shock. These elements appeared to act synergistically, since the intact promoter exhibited 30-fold more expression in stationary phase than the sum of that observed for each element acting independently. HOG1, which encodes a MAP kinase, has been implicated in control mediated by STREs. For GPH1, induction by osmotic shock depended on a functional HOG1 allele. In contrast, induction upon entry into stationary phase was only partially dependent on HOG1. Furthermore, the heat shock response, which can also be mediated by STREs, was independent of HOG1. These observations suggest that the GPH1 STREs respond to more than one pathway, only one of which requires HOG1.
Sunnarborg
S W
SW
Department of Biochemistry, Molecular Biology, and Biophysics, 6-155 Jackson Hall, University of Minnesota, Minneapolis, MN 55455, USA.
Miller
S P
SP
Unnikrishnan
I
I
LaPorte
D C
DC
D001483:Base Sequence; D003001:Cloning, Molecular; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics* / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000235:genetics* / Q000502:physiology; D006003:Glycogen / Q000378:metabolism; D024981:Glycogen Phosphorylase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009997:Osmotic Pressure; D012333:RNA, Messenger / Q000737:chemistry; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D016385:TATA Box / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis
D012333:RNA, Messenger; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D006003:Glycogen; D024981:Glycogen Phosphorylase; C088589:HOG1 protein, S cerevisiae; D020928:Mitogen-Activated Protein Kinases; D001616:beta-Galactosidase