Ultraviolet-B-mediated induction of protein-protein interactions in mammalian cells

Nat Commun. 2013:4:1779. doi: 10.1038/ncomms2800.

Abstract

Light-sensitive proteins are useful tools to control protein localization, activation and gene expression, but are currently limited to excitation with red or blue light. Here we report a novel optogenetic system based on the ultraviolet-B-dependent interaction of the Arabidopsis ultraviolet-B photoreceptor UVR8 with COP1 that can be performed in visible light background. We use this system to induce nuclear accumulation of cytoplasmic green fluorescent protein fused to UVR8 in cells expressing nuclear COP1, and to recruit a nucleoplasmic red fluorescent protein fused to COP1 to chromatin in cells expressing UVR8-H2B. We also show that ultraviolet-B-dependent interactions between DNA-binding and transcription activation domains result in a linear induction of gene expression. The UVR8-COP1 interactions in mammalian cells can be induced using subsecond pulses of ultraviolet-B light and last several hours. As UVR8 photoperception is based on intrinsic tryptophan residues, these interactions do not depend on the addition of an exogenous chromophore.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / metabolism
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Cell Nucleus / radiation effects
  • Chromatin / metabolism
  • DNA Damage
  • Green Fluorescent Proteins / metabolism
  • Histones / metabolism
  • Humans
  • Mammals / metabolism*
  • NF-kappa B / chemistry
  • NF-kappa B / metabolism
  • Protein Binding / radiation effects
  • Protein Structure, Tertiary
  • Protein Transport / radiation effects
  • Recombinant Fusion Proteins / metabolism
  • Time Factors
  • Transcription, Genetic / radiation effects
  • Ultraviolet Rays*

Substances

  • Arabidopsis Proteins
  • Chromatin
  • H2AX protein, human
  • Histones
  • NF-kappa B
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins