Transcriptional regulators of the Schizosaccharomyces pombe fbp1 gene include two redundant Tup1p-like corepressors and the CCAAT binding factor activation complex

Genetics. 2001 Mar;157(3):1205-15.

Abstract

The Schizosaccharomyces pombe fbp1 gene, which encodes fructose-1,6-bis-phosphatase, is transcriptionally repressed by glucose through the activation of the cAMP-dependent protein kinase A (PKA) and transcriptionally activated by glucose starvation through the activation of a mitogen-activated protein kinase (MAPK). To identify transcriptional regulators acting downstream from or in parallel to PKA, we screened an adh-driven cDNA plasmid library for genes that increase fbp1 transcription in a strain with elevated PKA activity. Two such clones express amino-terminally truncated forms of the S. pombe tup12 protein that resembles the Saccharomyces cerevisiae Tup1p global corepressor. These clones appear to act as dominant negative alleles. Deletion of both tup12 and the closely related tup11 gene causes a 100-fold increase in fbp1-lacZ expression, indicating that tup11 and tup12 are redundant negative regulators of fbp1 transcription. In strains lacking tup11 and tup12, the atf1-pcr1 transcriptional activator continues to play a central role in fbp1-lacZ expression; however, spc1 MAPK phosphorylation of atf1 is no longer essential for its activation. We discuss possible models for the role of tup11- and tup12-mediated repression with respect to signaling from the MAPK and PKA pathways. A third clone identified in our screen expresses the php5 protein subunit of the CCAAT-binding factor (CBF). Deletion of php5 reduces fbp1 expression under both repressed and derepressed conditions. The CBF appears to act in parallel to atf1-pcr1, although it is unclear whether or not CBF activity is regulated by PKA.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • CCAAT-Binding Factor / genetics*
  • Cloning, Molecular
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • DNA, Complementary / metabolism
  • Fructose-Bisphosphatase / genetics*
  • Fungal Proteins / genetics*
  • Gene Deletion
  • Gene Expression Regulation, Fungal*
  • Gene Library
  • Genotype
  • MAP Kinase Signaling System
  • Molecular Sequence Data
  • Mutation
  • Nuclear Proteins*
  • Phenotype
  • Phosphorylation
  • Plasmids / genetics
  • Promoter Regions, Genetic
  • Repressor Proteins*
  • Saccharomyces cerevisiae Proteins*
  • Schizosaccharomyces / genetics*
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • Temperature
  • Transcription, Genetic*
  • Transcriptional Activation
  • beta-Galactosidase / metabolism

Substances

  • CCAAT-Binding Factor
  • DNA, Complementary
  • Fungal Proteins
  • Nuclear Proteins
  • Repressor Proteins
  • Saccharomyces cerevisiae Proteins
  • TUP1 protein, S cerevisiae
  • Cyclic AMP-Dependent Protein Kinases
  • Fructose-Bisphosphatase
  • beta-Galactosidase