title: Suppressing the Neurospora crassa circadian clock while maintaining light responsiveness in continuous stirred tank reactors.
abstract: Neurospora crassa has been utilized as a model organism for studying biological, regulatory, and circadian rhythms for over 50 years. These circadian cycles are driven at the molecular level by gene transcription events to prepare for environmental changes. N. crassa is typically found on woody biomass and is commonly studied on agar-containing medium which mimics its natural environment. We report a novel method for disrupting circadian gene transcription while maintaining light responsiveness in N. crassa when held in a steady metabolic state using bioreactors. The arrhythmic transcription of core circadian genes and downstream clock-controlled genes was observed in constant darkness (DD) as determined by reverse transcription-quantitative PCR (RT-qPCR). Nearly all core circadian clock genes were up-regulated upon exposure to light during 11hr light/dark cycle experiments under identical conditions. Our results demonstrate that the natural timing of the robust circadian clock in N. crassa can be disrupted in the dark when maintained in a consistent metabolic state. Thus, these data lead to a path for the production of industrial scale enzymes in the model system, N. crassa, by removing the endogenous negative feedback regulation by the circadian oscillator.
authors.affiliation: Chemistry Division, US Naval Research Laboratory, 4555 Overlook Ave., SW., Washington, DC, 20375, USA.
authors.forename: Allison L
authors.initials: AL
authors.lastname: Cockrell
authors.affiliation: Chemistry Division, US Naval Research Laboratory, 4555 Overlook Ave., SW., Washington, DC, 20375, USA.
authors.forename: Russell K
authors.initials: RK
authors.lastname: Pirlo
authors.affiliation: Nova Research Inc., 1900 Elkin St., Suite 230, Alexandria, VA, 22308, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Babson
authors.affiliation: Chemistry Division, US Naval Research Laboratory, 4555 Overlook Ave., SW., Washington, DC, 20375, USA.
authors.forename: Kathleen D
authors.initials: KD
authors.lastname: Cusick
authors.affiliation: Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, 4555 Overlook Ave., SW., Washington, DC, 20375, USA.
authors.forename: Carissa M
authors.initials: CM
authors.lastname: Soto
authors.affiliation: Nova Research Inc., 1900 Elkin St., Suite 230, Alexandria, VA, 22308, USA.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Petersen
authors.affiliation: Howard University, Washington, DC. 20057.
authors.forename: Miah J
authors.initials: MJ
authors.lastname: Davis
authors.affiliation: Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, OH 45267, USA.
authors.forename: Christian I
authors.initials: CI
authors.lastname: Hong
authors.affiliation: Department of Biology, Rutgers University, Camden, NJ, 08102, USA.
authors.forename: Kwangwon
authors.initials: K
authors.lastname: Lee
authors.affiliation: Chemistry Division, US Naval Research Laboratory, 4555 Overlook Ave., SW., Washington, DC, 20375, USA.
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Fitzgerald
authors.affiliation: Chemistry Division, US Naval Research Laboratory, 4555 Overlook Ave., SW., Washington, DC, 20375, USA.
authors.forename: Justin C
authors.initials: JC
authors.lastname: Biffinger
mesh_terms: D057906:Circadian Clocks / Q000235:genetics; D002940:Circadian Rhythm / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000528:radiation effects; D008027:Light; D009492:Neurospora crassa / Q000502:physiology* / Q000528:radiation effects*; D017440:Photoperiod; D014158:Transcription, Genetic
chemical_list: D005656:Fungal Proteins