Characterization of VOS-1 transcription factor in response to different stress conditions and the connection with glycogen accumulation in Neurospora crassa

• Grant number: 14/24627-5
• Support type: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): March 01, 2015
• Effective date (End): February 29, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Maria Celia Bertolini
• Grantee: Amanda Ventura Campos Araujo
• Home Institution: Instituto de Química (IQ). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

The genome of the fungus Neurospora crassa was sequenced in 2003 (Galagan et al.), revealing the existence of about 10,000 genes encoding proteins. Recent analysis showed that only 40% of the identified genes correspond to known proteins (Wang et al., 2011). N. crassa is a model organism used in studies of gene expression, development and cell differentiation, circadian clock and genome defense (Perkins and Davis, 2000). After the genome sequencing, the construction of knockout strains in each gene was started and the collection was available to the scientific community. In our laboratory, a screening using mutant strains in genes encoding transcription factors was performed to analyze the glycogen accumulation after growing under normal temperature (30ºC) and after heat shock (30ºC’45ºC). Some mutant strains showed changes in the glycogen accumulation profile when compared to the wild-type strain (Gonçalves et al., 2011, Boni et al., in preparation). Thus, the transcription factors missing in the mutant strains were considered as proteins likely involved in the regulation of glycogen metabolism. Among these transcription factors, the protein VOS-1 protein was shown to regulate glycogen levels in conidia and in mycelia during vegetative growth (Boni et al., in preparation). In addition, the protein was shown to regulate glycogen levels during circadian clock experiments by controlling the expression of the genes gsn and gpn (Virgilio et al., in preparation). Morphological assays and results of RNA-seq and ChIP-seq experiments showed that VOS-1 likely regulates genes involved in stress response. This project aims to perform the characterization of VOS-1 under different stressing conditions by analyzing the growth of the mutant strain, the cellular localization of the protein, and also to investigate the regulation of glycogen metabolism by VOS-1 under the same conditions.