Estudo do fluxo de prótons em Saccharomyces cerevisiae sob estresse etanólico
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Data
2022-05-02
Autores
Pinto, Camila Moreira
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Universidade Estadual Paulista (Unesp)
Resumo
Saccharomyces cerevisiae é amplamente utilizada na produção de bioetanol e as elevadas concentrações de etanol durante a fermentação representam um dos estressores mais comuns que afetam inicialmente a membrana citoplasmática. Entretanto, o conhecimento acerca da dinâmica dos compartimentos da membrana plasmática em S. cerevisiae sob estresse etanólico é incipiente. Aqui, objetivou-se buscar por características que contribuam para a melhor compreensão da dinâmica dos compartimentos eisossoma e MCP em resposta ao estresse etanólico em S. cerevisiae. Durante 240 min, células da linhagem BY4741 foram expostas a diferentes concentrações de etanol e analisou-se a acidificação do meio extracelular, taxa de crescimento, sublocalização celular de proteínas e análises da expressão gênica e proteica. Os resultados mostraram que o etanol tem influência na interação entre os compartimentos MCP e eisossoma, afetando diretamente o fluxo de prótons entre a célula e o meio externo e o nível de interferência é norteado pelo percentual de etanol. Adicionalmente, a redução da Pma1p (MCP) em células sob estresse por etanol corrobora a baixa atividade de fluxo de prótons. O aumento de eisossomas juntamente com o aumento de Can1p e Xrn1p sugerem um papel protetor e regulador do eisossoma durante o estresse por etanol. Por fim, demonstramos o aumento de grânulos de estresse vinculado ao aumento de eisossomas durante o estresse por etanol. Em suma, reportamos algumas características-chave para a compreensão da dinâmica dos compartimentos eisossoma e MCP em resposta estresse etanólico, evidenciamos a importância da interação entre os compartimentos de membrana plasmática com vias regulatórias responsáveis pelo homeostase iônica intracelular e degradação de mRNA e sugerimos que eisossomas podem representar uma importante resposta celular adaptativa ao estresse por etanol.
Saccharomyces cerevisiae is widely used in the bioethanol production and the high ethanol concentrations during fermentation represent one of the most common stressors that initially affect the cytoplasmic membrane. However, the knowledge about dynamics of plasma membrane compartments in S. cerevisiae under ethanolic stress is incipient. Here, the objective was to search for characteristics that contribute to a better understanding of the dynamics of eisosome and MCP compartments in response to ethanolic stress in S. cerevisiae. During 240 min, BY4741 cells were exposed to different ethanol concentrations and the acidification of extracellular environment, growth rate, cellular protein sublocalization and gene and protein expression analysis were analyzed. The results showed that ethanol has an influence on the interaction between MCP and eisosome compartments, directly affecting the proton flow between cell and external environment, and the interference level is guided by the ethanol percentage. Additionally, the Pma1p (MCP) reduction in cells under ethanol stress corroborates with the proton flux activity low. The increase in eisosomes together with the increase in Can1p and Xrn1p suggest a protective and regulatory role of the eisosome during ethanol stress. Finally, we demonstrate the increase in stress granules linked to the increase in eisosomes during ethanol stress. In summary, we report some key-features for understanding the dynamics of eisosome and MCP compartments in response to ethanolic stress, highlight the importance of interaction between plasma membrane compartments with regulatory pathways responsible for intracellular ionic homeostasis and mRNA degradation and suggest that eisosomes may represent an important adaptive cellular response to ethanol stress.
Saccharomyces cerevisiae is widely used in the bioethanol production and the high ethanol concentrations during fermentation represent one of the most common stressors that initially affect the cytoplasmic membrane. However, the knowledge about dynamics of plasma membrane compartments in S. cerevisiae under ethanolic stress is incipient. Here, the objective was to search for characteristics that contribute to a better understanding of the dynamics of eisosome and MCP compartments in response to ethanolic stress in S. cerevisiae. During 240 min, BY4741 cells were exposed to different ethanol concentrations and the acidification of extracellular environment, growth rate, cellular protein sublocalization and gene and protein expression analysis were analyzed. The results showed that ethanol has an influence on the interaction between MCP and eisosome compartments, directly affecting the proton flow between cell and external environment, and the interference level is guided by the ethanol percentage. Additionally, the Pma1p (MCP) reduction in cells under ethanol stress corroborates with the proton flux activity low. The increase in eisosomes together with the increase in Can1p and Xrn1p suggest a protective and regulatory role of the eisosome during ethanol stress. Finally, we demonstrate the increase in stress granules linked to the increase in eisosomes during ethanol stress. In summary, we report some key-features for understanding the dynamics of eisosome and MCP compartments in response to ethanolic stress, highlight the importance of interaction between plasma membrane compartments with regulatory pathways responsible for intracellular ionic homeostasis and mRNA degradation and suggest that eisosomes may represent an important adaptive cellular response to ethanol stress.
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Palavras-chave
Estresse etanólico, Eisossoma, Fluxo de prótons, Saccharomyces cerevisiae, Ethanolic stress, Eisosome, Proton flux, Etanol, Leveduras (Fungos)