Abstract
Genome sequencing allows to perform regulatory studies in a deep way. In living cells genomes are not simply made of DNA and proteins, but rather of a large amount of regulatory proteins that dynamically interact with DNA and modulate gene expression. Our laboratory has been using the filamentous fungus Neurospora crassa as a model organism to study the molecular mechanisms that are involved in the glycogen metabolism regulation. Previous results showed that the metabolism of this carbohydrate is connected with signaling pathways participating in different cellular processes, which suggests the importance of the energy produced by this metabolic pathway to the biological processes. In a previous project (Proc. 08/57566-8), some transcription factors, identified as proteins regulating the glycogen metabolism in N. crassa, were functionally characterized and many of them are being studied at the present moment. In a similar way, a collection of mutant strains in protein kinases was screened and many kinases were identified as likely regulating the glycogen metabolism. This project aims to continue the characterization studies focusing in the functional and structural aspects of transcription factors and protein kinases not yet characterized in N. crassa. Considering the previous results the main objectives of this project are: 1) to investigate how some transcription factors act in the glycogen metabolism regulation, such as FLBC, a transcription factor involved in asexual development, NIT2, a transcription factor that regulates nitrogen metabolism in N. crassa and transcriptional factors annotated as hipothetical proteins. Different technical approaches will be used to achieve this goal, 2) to investigate how some transcription factor are transported to the nucleus using co-crystallization assays with the protein importin of N. crassa and Isotermal Titration Calorimetry (ITC), 3) to functionally characterize protein kinases previously identified as regulators of glycogen metabolism, 4) to characterize the connection between the circadian clock and the energy of the glycogen metabolism. In this project we expect to bring new and relevant information concerning the functional genomic of the fungus, more specifically in relation to the regulatory network existent in N. crassa. (AU)
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