The utilization of plant biomass by filamentous fungi requires the orchestrated production and secretion of hydrolytic enzymes, followed by transport and intracellular utilization of plant cell wall components. In industrial settings, hydrolytic enzyme production is important for the production of 2nd generation biofuels and specialty chemicals from lignocellulose. However, rational engineering of filamentous fungi for enzyme production is hampered by a dearth of integrated metabolic information that precludes the development of predictive models for spatiotemporal dynamics of plant biomass degradation. We therefore propose to characterize the metabolic network of the model cellulolytic filamentous fungus, Neurospora crassa. The primary objectives of this proposal are to assess phosphoproteomic and metabolomic responses when N. crassa is exposed to, and subsequently utilizes, plant biomass. First, alterations of the phosphoproteome that are most likely associated with sensing of plant biomass in the environment will be determined. Second, the steady state phosphoproteome and metabolome will be determined, which are predicted to be associated with the integration of fungal metabolism and utilization of plant biomass. The final objective of this proposal is to interrogate the metabolic integration of the utilization of carbon (as cellulose) with nitrogen, phosphorus and sulfur. In nature, these elements are often limiting, suggesting that these nutrient pathways must be highly integrated. Our ultimate goal is to develop a high-resolution, metabolic network for a filamentous fungus by the integration of proteomic, metabolomic and transcriptomic data and thereby generate a predictive and dynamic model for plant biomass deconstruction by filamentous fungi.