Role of the N-terminal region of Rap1p in the transcriptional activation of glycolytic genes in Saccharomyces cerevisiae

Yeast. 2004 Jul 30;21(10):851-66. doi: 10.1002/yea.1123.

Abstract

In the yeast two-hybrid system, the N-terminal region of Rap1p was shown to interact with Gcr1p and Gcr2p. Disruption of gcr1 and/or gcr2 in the two-hybrid reporter strain demonstrated that the interaction with Gcr1p does not require Gcr2p, whereas the interaction with Gcr2p is mediated through Gcr1p. Deletion of the N-terminal region of Rap1p alone did not show a growth phenotype, but a growth defect was observed when this mutation was combined with a gcr2 deletion. The poor growth of the gcr1 null mutant was not affected further by the N-terminal deletion of Rap1p, but the growth of gcr1 strains with mutations in the DNA binding region of Gcr1p was affected by the removal of the N-terminal region of Rap1p. These results suggest that one function of the N-terminal region of Rap1p, presumably the BRCT domain, is to facilitate the binding of Gcr1p to the promoter by a protein-protein interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • DNA, Fungal / genetics
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Gene Expression Regulation, Fungal / genetics
  • Gene Expression Regulation, Fungal / physiology*
  • Mutagenesis
  • RNA, Fungal / chemistry
  • RNA, Fungal / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Shelterin Complex
  • Telomere-Binding Proteins / genetics
  • Telomere-Binding Proteins / metabolism*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcriptional Activation / genetics
  • Transcriptional Activation / physiology*
  • Two-Hybrid System Techniques

Substances

  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • GCR1 protein, S cerevisiae
  • GCR2 protein, S cerevisiae
  • RAP1 protein, S cerevisiae
  • RNA, Fungal
  • Saccharomyces cerevisiae Proteins
  • Shelterin Complex
  • Telomere-Binding Proteins
  • Trans-Activators
  • Transcription Factors