A common docking site for response regulators on the yeast phosphorelay protein YPD1

Biochim Biophys Acta. 2005 May 15;1748(2):138-45. doi: 10.1016/j.bbapap.2004.12.009. Epub 2005 Jan 7.

Abstract

In Saccharomyces cerevisiae, a multi-component phosphorelay signal transduction pathway mediates cellular responses to environmental stress. A histidine-containing phosphotransfer protein, YPD1, represents a bifurcation point between the SLN1-YPD1-SSK1 pathway responsible for osmotic stress responses and the SLN1-YPD1-SKN7 pathway involved in cell wall biosynthesis and cell cycle control. The phosphorelay protein YPD1 must physically interact with and transfer phosphoryl groups between three homologous response regulator domains, designated SLN1-R1, SSK1-R2, and SKN7-R3. In this comparative study, the molecular basis of interaction was examined between YPD1 and each of the three response regulator domains utilizing alanine scanning mutagenesis combined with a yeast two-hybrid assay. Results from the yeast two-hybrid assay indicate that all three response regulator domains bind to a common area, largely hydrophobic in nature, on the surface of YPD1. We postulate that other YPD1 surface residues surrounding this common docking site are involved in making specific interactions with one or more of the response regulator domains.

Publication types

  • Comparative Study

MeSH terms

  • Alanine / chemistry
  • Binding Sites
  • Cell Cycle
  • Cell Wall / metabolism
  • Crystallography, X-Ray
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism
  • Gene Library
  • Genes, Regulator
  • Genetic Vectors
  • Histidine / chemistry
  • Intracellular Signaling Peptides and Proteins
  • Models, Molecular
  • Mutagenesis
  • Mutation
  • Osmosis
  • Plasmids / metabolism
  • Protein Binding
  • Protein Conformation
  • Protein Kinases / metabolism
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / physiology*
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Transcription Factors / metabolism
  • Two-Hybrid System Techniques

Substances

  • DNA-Binding Proteins
  • Intracellular Signaling Peptides and Proteins
  • SKN7 protein, S cerevisiae
  • SSK1 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Histidine
  • Protein Kinases
  • YPD1 protein, S cerevisiae
  • SLN1 protein, S cerevisiae
  • Alanine