title: The SEB-1 Transcription Factor Binds to the STRE Motif in Neurospora crassa and Regulates a Variety of Cellular Processes Including the Stress Response and Reserve Carbohydrate Metabolism.
abstract: When exposed to stress conditions, all cells induce mechanisms resulting in an attempt to adapt to stress that involve proteins which, once activated, trigger cell responses by modulating specific signaling pathways. In this work, using a combination of pulldown assays and mass spectrometry analyses, we identified the Neurospora crassa SEB-1 transcription factor that binds to the Stress Response Element (STRE) under heat stress. Orthologs of SEB-1 have been functionally characterized in a few filamentous fungi as being involved in stress responses; however, the molecular mechanisms mediated by this transcription factor may not be conserved. Here, we provide evidences for the involvement of N. crassa SEB-1 in multiple cellular processes, including response to heat, as well as osmotic and oxidative stress. The Δseb-1 strain displayed reduced growth under these conditions, and genes encoding stress-responsive proteins were differentially regulated in the Δseb-1 strain grown under the same conditions. In addition, the SEB-1-GFP protein translocated from the cytosol to the nucleus under heat, osmotic, and oxidative stress conditions. SEB-1 also regulates the metabolism of the reserve carbohydrates glycogen and trehalose under heat stress, suggesting an interconnection between metabolism control and this environmental condition. We demonstrated that SEB-1 binds in vivo to the promoters of genes encoding glycogen metabolism enzymes and regulates their expression. A genome-wide transcriptional profile of the Δseb-1 strain under heat stress was determined by RNA-seq, and a broad range of cellular processes was identified that suggests a role for SEB-1 as a protein interconnecting these mechanisms.
authors.affiliation: Departamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista (UNESP), 14800-060, Araraquara, São Paulo, Brazil.
authors.forename: Fernanda Zanolli
authors.initials: FZ
authors.lastname: Freitas
authors.affiliation: Departamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista (UNESP), 14800-060, Araraquara, São Paulo, Brazil.
authors.forename: Stela
authors.initials: S
authors.lastname: Virgilio
authors.affiliation: Departamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista (UNESP), 14800-060, Araraquara, São Paulo, Brazil.
authors.forename: Fernanda Barbosa
authors.initials: FB
authors.lastname: Cupertino
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley, California, 94720-3102.
authors.forename: David John
authors.initials: DJ
authors.lastname: Kowbel
authors.affiliation: Departamento de Química Orgânica, Instituto de Química, Universidade Estadual de Campinas (UNICAMP), 13803-862, Campinas, São Paulo, Brazil.
authors.forename: Mariana
authors.initials: M
authors.lastname: Fioramonte
authors.affiliation: Departamento de Química Orgânica, Instituto de Química, Universidade Estadual de Campinas (UNICAMP), 13803-862, Campinas, São Paulo, Brazil.
authors.forename: Fabio Cesar
authors.initials: FC
authors.lastname: Gozzo
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley, California, 94720-3102.
authors.forename: N Louise
authors.initials: NL
authors.lastname: Glass
authors.affiliation: Departamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista (UNESP), 14800-060, Araraquara, São Paulo, Brazil mcbertol@iq.unesp.br.
authors.forename: Maria Célia
authors.initials: MC
authors.lastname: Bertolini
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D050260:Carbohydrate Metabolism; D047369:Chromatin Immunoprecipitation; D004777:Environment; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D059014:High-Throughput Nucleotide Sequencing; D009492:Neurospora crassa / Q000235:genetics* / Q000378:metabolism*; D059372:Nucleotide Motifs; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020218:Response Elements; D013312:Stress, Physiological / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D014157:Transcription Factors
keywords: ChIP-qPCR; Neurospora crassa; RNA-seq; SEB-1; stress response