The Schizosaccharomyces pombe MBF complex requires heterodimerization for entry into S phase

Mol Cell Biol. 1995 May;15(5):2589-99. doi: 10.1128/MCB.15.5.2589.

Abstract

In Schizosaccharomyces pombe, MBF is a DNA-binding complex suspected to activate the transcription of genes necessary for entry into S phase. The MBF complex contains both p85cdc10 and p72res1/sct1. To obtain a better understanding of how the MBF complex regulates gene expression at the G1/S transition, we have performed a genetic analysis of p72res1. We determined that p72res1 can bind specifically to the cdc22 promoter, when analyzed by gel mobility shift assay, and that the N-terminal 157 amino acids of p72res1 are sufficient for this specific binding. When overexpressed in vivo, a fragment of p72res1 containing this DNA-binding domain could rescue a strain carrying a temperature-sensitive cdc10 allele at the restrictive temperature as well as a strain with a cdc10 null allele. We also determined that the C-terminal region of p72res1 is necessary and sufficient for binding to p85cdc10. Overexpression of the cdc10-binding domain of p72res1 leads to a G1 arrest with a cdc phenotype and a decrease on MBF activity. Overexpression of full-length p72res1 also leads to a growth arrest that can be rescued by overexpression of p85cdc10. These results imply that the MBF activity in vivo is dependent on the interaction of p85cdc10 with p72res1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites / genetics
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • DNA Primers / genetics
  • DNA, Fungal / genetics
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Molecular Sequence Data
  • Mutation
  • Protein Conformation
  • S Phase
  • Schizosaccharomyces / cytology
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins*
  • Sequence Deletion
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • Cell Cycle Proteins
  • DNA Primers
  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • Schizosaccharomyces pombe Proteins
  • Transcription Factors
  • cdc10 protein, S pombe
  • res1 protein, S pombe