title: Light-dependent and circadian transcription dynamics in vivo recorded with a destabilized luciferase reporter in Neurospora.
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.
authors.affiliation: University of Heidelberg Biochemistry Center (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
authors.forename: François
authors.initials: F
authors.lastname: Cesbron
authors.affiliation: University of Heidelberg Biochemistry Center (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Brunner
authors.affiliation: University of Heidelberg Biochemistry Center (BZH), Im Neuenheimer Feld, Heidelberg, Germany.
authors.forename: Axel C R
authors.initials: AC
authors.lastname: Diernfellner
mesh_terms: D000595:Amino Acid Sequence; D002940:Circadian Rhythm / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D008027:Light; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009491:Neurospora / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: C087405:FRQ protein, Neurospora crassa; D005656:Fungal Proteins; D008156:Luciferases