A LOV2 domain-based optogenetic tool to control protein degradation and cellular function

Chem Biol. 2013 Apr 18;20(4):619-26. doi: 10.1016/j.chembiol.2013.03.005.

Abstract

Light perception is indispensable for plants to respond adequately to external cues and is linked to proteolysis of key transcriptional regulators. To provide synthetic light control of protein stability, we developed a generic photosensitive degron (psd) module combining the light-reactive LOV2 domain of Arabidopsis thaliana phot1 with the murine ornithine decarboxylase-like degradation sequence cODC1. Functionality of the psd module was demonstrated in the model organism Saccharomyces cerevisiae. Generation of conditional mutants, light regulation of cyclin-dependent kinase activity, light-based patterning of cell growth, and yeast photography exemplified its versatility. In silico modeling of psd module behavior increased understanding of its characteristics. This engineered degron module transfers the principle of light-regulated degradation to nonplant organisms. It will be highly beneficial to control protein levels in biotechnological or biomedical applications and offers the potential to render a plethora of biological processes light-switchable.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / chemistry
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Cell Cycle Checkpoints / drug effects
  • Light*
  • Mice
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Ornithine Decarboxylase / chemistry
  • Ornithine Decarboxylase / genetics
  • Ornithine Decarboxylase / metabolism*
  • Phosphoproteins / chemistry
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Protein Stability
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases
  • Proteolysis
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / pharmacology
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / metabolism

Substances

  • Arabidopsis Proteins
  • Phosphoproteins
  • Recombinant Fusion Proteins
  • NPH1 protein, Arabidopsis
  • Protein-Serine-Threonine Kinases
  • Ornithine Decarboxylase