Localization and light-dependent phosphorylation of white collar 1 and 2, the two central components of blue light signaling in Neurospora crassa

Eur J Biochem. 2000 Jan;267(2):414-22. doi: 10.1046/j.1432-1327.2000.01016.x.

Abstract

In Neurospora crassa only two white collar (wc) mutants, wc-1 and wc-2, have been described that seem to be insensitive to light. The pleiotropic phenotypes of these mutants suggest that they represent two central components of blue light signal transduction. The WC proteins have several characteristics of transcription factors consistent with an involvement in transcriptional control of light-regulated genes. Here, we present a biochemical analysis of WC1 and WC2 polypeptides in N. crassa. Using specific antisera against WC1 and WC2, respectively, the subcellular localization of the WC polypeptides was investigated. The WC1 protein was localized exclusively in the nucleus, whereas WC2 was detected in both the nuclear and cytoplasmic fractions. The nuclear localization of WC1 and WC2 was shown to be independent of light and dimerization between the two proteins. In addition, WC1 and WC2 are phosphorylated in response to light. The phosphorylation of WC1 and WC2 was dependent on functional WC1 and WC2 proteins, respectively, which clearly indicated a correlation between the light-dependent phosphorylation and the function of WC1 and WC2 in blue light signaling. However, the light-specific phosphorylation of the WC proteins revealed different kinetics. The phosphorylation of WC1 was transient whereas the WC2 phosphorylation was shown to be stable under constant light conditions. The analysis of the light-dependent phosphorylation of WC1 and WC2 in wc-2 and wc-1 mutants revealed an epistatic relationship for WC1 and WC2 with WC2 acting downstream of WC1 in the signal transduction pathway of blue light.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / metabolism
  • Chromatography, Affinity
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / immunology
  • DNA-Binding Proteins / metabolism*
  • Fungal Proteins / immunology
  • Fungal Proteins / metabolism*
  • Immune Sera
  • Immunoglobulin G / isolation & purification
  • Light
  • Mutation
  • Neurospora crassa / physiology*
  • Nuclear Proteins / immunology
  • Nuclear Proteins / metabolism
  • Photophosphorylation / physiology*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Signal Transduction*
  • Transcription Factors / genetics
  • Transcription Factors / immunology
  • Transcription Factors / metabolism*

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • Immune Sera
  • Immunoglobulin G
  • Nuclear Proteins
  • Recombinant Proteins
  • Transcription Factors
  • wc-1 protein, Neurospora crassa
  • white collar 2 protein, Neurospora