Effects of Time of Day on the Neurospora Metabolome
EMSL Project ID
47818
Abstract
Biofuel production using cellulases extracted from fungi has the potential to be a major resource to replace fossil fuels and regulating the synthesis of these and other enzymes is crucial for the economics of biofuel production from lignocellulose. Some of the best inducers of plant cell wall-degrading enzymes are filamentous fungi. However, environmental controls on cellular metabolism are known to modulate the amount of cellulases that can be produced in fungi. The filamentous fungus Neurospora is commonly used in the production of cellulases for biofuel manufacture and it is also the principal fungal model system for the study of the effects of light and circadian clocks. The circadian clock plays a large role in the regulation of the Neurospora genome, with far reaching effects, including many processes essential for growth and development, specifically including metabolism. Although the dominant general role of the clock in metabolic regulation is appreciated, the degree to which the clock controls specific aspects of metabolism is not currently understood in any organism.
This project seeks to identify the link between time of day and cellulase levels in an effort to better understand their regulation, and moreover to improve the production of these important components for energy production. We know that one of the proteins regulated by the clock, CSP-1, is predominantly involved in regulating metabolism, and could directly impact cellulase production. Removal of CSP-1by targeted deletion provides an opportunity to eliminate circadian regulation of metabolism, potentially increasing and stabilizing cellulase production.
To understand the role that the circadian clock plays in regulating cellular metabolism, and more specifically in the expression of cellulases used in biofuel production, we will determine the metabolic profile of two strains of Neurospora crassa, both a wild type strain and a deletion mutation (?csp-1) in which the link between metabolism and the clock is crippled. We will rely on the technical abilities of the EMSL to provide a thorough mass spectrometry (MS) analysis of the Neurospora metabolome over a circadian cycle and to process the large amount of raw data that will be procured. We will then use commonly applied informatic and statistical analyses to determine which metabolites are rhythmic and the degree to which the loss of CSP-1 affects the circadian clock. After analysis of these data, we will track known cellulases more directly via comparative MS to determine conditions under which optimal levels of cellulases are created, whether at a specific time of day or in a strain with no clock regulation on metabolism. Again, the resources of the EMSL will be essential in completing this goal, as the DOE prosecute the proteomics using iTRAQ. As with the metabolomic analysis, the EMSL will analyze the raw data, which will then be sifted using informatics and statistical analysis commonly used within the circadian field. We hypothesize that these data will provide insights into whether and how to decouple metabolism from the clock with the goal of increasing the production of cellulases needed in biofuel manufacture.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2013-10-01
End Date
2014-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
Team Members
Related Publications
Hurley J.M., A. Dasgupta, J.M. Emerson, X. Zhou, C. Ringelberg, N. Knabe, and A. Lipzen, et al. 2014. "Analysis of Clock-regulated Genes in Neurospora Reveals Widespread Posttranscriptional Control of Metabolic Potential." Proceedings of the National Academy of Sciences (PNAS). 111, no. 48:16995–17002. doi:10.1073/pnas.1418963111
Hurley J.M., M.S. Jankowski, H. De Los Santos, A.M. Crowell, S. Fordyce, J.D. Zucker, and N. Kumar, et al. 2018. "Circadian proteomic analysis uncovers mechanisms of post-transcriptional regulation in metabolic pathways." Cell Systems 7, no. 6:613-626. PNNL-SA-138421. doi:10.1016/j.cels.2018.10.014