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G3 (Bethesda) ; 6(5): 1327-43, 2016 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-26994287

RESUMO

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.


Assuntos
Sítios de Ligação , Metabolismo dos Carboidratos , Neurospora crassa/genética , Neurospora crassa/metabolismo , Motivos de Nucleotídeos , Elementos de Resposta , Estresse Fisiológico , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Imunoprecipitação da Cromatina , Meio Ambiente , Deleção de Genes , Regulação Fúngica da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Fenótipo , Regiões Promotoras Genéticas , Ligação Proteica , Estresse Fisiológico/genética
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