Light-dependent and circadian transcription dynamics in vivo recorded with a destabilized luciferase reporter in Neurospora

PLoS One. 2013 Dec 31;8(12):e83660. doi: 10.1371/journal.pone.0083660. eCollection 2013.

Abstract

We show that firefly luciferase is a stable protein when expressed at 25 °C in Neurospora, which limits its use as transcription reporter. We created a short-lived luciferase by fusing a PEST signal to its C-terminus (LUC-PEST) and applied the LUC-PEST reporter system to record in vivo transcription dynamics associated with the Neurospora circadian clock and its blue-light photosensory system over the course of several days. We show that the tool is suitable to faithfully monitor rapid, but also subtle changes in transcription in a medium to high throughput format.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Circadian Rhythm / genetics*
  • Fungal Proteins / genetics*
  • Light*
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Luminescent Measurements
  • Molecular Sequence Data
  • Neurospora / genetics*
  • Neurospora / metabolism
  • Polymerase Chain Reaction
  • Transcription, Genetic*

Substances

  • FRQ protein, Neurospora crassa
  • Fungal Proteins
  • Luciferases

Grants and funding

This work was supported by grants from the Deutsche Forschungsgemeinschaft: DFG DI 1874/1-1 to AD (http://www.dfg.de). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.