title: Accounting for strain-specific differences during RTG target gene regulation in Saccharomyces cerevisiae.
abstract: Mitochondrial dysfunction results in the expression, via the retrograde response pathway, of a concise set of genes (RTG target genes) that encode enzymes involved in the anapleurotic production of alpha-ketoglutarate. Inhibiting the rapamycin-sensitive TOR kinases, important regulators of cell growth, similarly results in RTG target gene expression under rich nutrient conditions. Retrograde and TOR-dependent regulation of RTG target genes requires a number of shared components, including the heterodimeric bZip/HLH transcription factors Rtg1p and Rtg3p, as well as their upstream regulator Mks1p. Two unresolved discrepancies exist with regard to the mechanism of RTG target gene control: (1) deletion of MKS1 results in constitutive expression of RTG target genes in most but not all strain backgrounds; and (2) RTG target gene expression has been correlated with both decreased as well as increased Rtg3p phosphorylation. Here we have addressed both of these issues. First, we demonstrate that the mks1 deletion strain used in a previous study by Shamji and coworkers contains a nonsense mutation within codon Ser 231 in RTG3 that likely accounts for the inactivity of the RTG system in this strain. Second, we confirm results by Butow and coworkers that Rtg3p is dephosphorylated as a primary response to induction of the pathway. Hyper-phosphorylation of this protein appears to be a secondary consequence of rapamycin treatment and is influenced both by strain background as well as by specific supplied nutrients. That hyper-phosphorylation of Rtg3p is also caused by heat shock suggests that it may reflect a more generalized response to cell stress. Together these results contribute toward a uniform view of RTG target gene regulation.
authors.affiliation: Section of Molecular and Cellular Biology & Center for Genetics and Development, College of Biological Sciences, University of California, Davis, CA 95616, USA.
authors.forename: Ivanka
authors.initials: I
authors.lastname: Dilova
authors.forename: Ted
authors.initials: T
authors.lastname: Powers
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media; D015966:Gene Expression Regulation, Fungal; D009154:Mutation; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000145:classification* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D020123:Sirolimus / Q000494:pharmacology; D013045:Species Specificity; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D003470:Culture Media; C080921:MKS1 protein, S cerevisiae; C104438:RTG3 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D017346:Protein Serine-Threonine Kinases; C500749:target of rapamycin protein, S cerevisiae; D020123:Sirolimus