Investigação do envolvimento de ZapA e ZapB de Xanthomonas citri subsp. citri na divisão celular e segregação cromossômica
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Data
2021-05-31
Autores
Hypolito, Giovane Böerner
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Universidade Estadual Paulista (Unesp)
Resumo
A bactéria Xanthomonas citri subsp citri (X. citri) é um fitopatógeno que ataca todas as variedades comerciais de citros, causando impacto significativo para a produção mundial de laranjas. A doença causada por esta bactéria é conhecida como cancro cítrico, que produz lesões corticosas salientes nos frutos, diminuindo a produtividade e qualidade dos mesmos. Até o presente, não há cura para a cancro cítrico, sendo o manejo da doença uma das formas de controle utilizada para se evitar grandes perdas econômicas. Alternativamente, o estudo da biologia do patógeno possibilita o desenvolvimento de estratégias de controle utilizando compostos antibacterianos capazes de inibir alvos/processos vitais tais como a divisão celular. A maquinaria responsável pela divisão bacteriana é formada principalmente pela proteína FtsZ, uma tubulina ancestral que forma o anel-Z no centro das células em divisão. O anel-Z recruta diversas proteínas que atuarão no processo de divisão, bem como outras com função acessória na modulação de sua montagem. Caracterizar a função destas proteínas constitui uma forma de ampliar nossos alvos para o desenvolvimento de novos compostos inibidores de divisão celular, forma esta que constitui uma alternativa de controle do cancro cítrico. Neste trabalho, caracterizamos duas proteínas acessórias, ZapA e ZapB, codificadas por este patógeno. Investigamos suas funções avaliando o fenótipo de mutante de X. citri com deleção de zapB (XAC_RS17260) e zapA (XAC_RS17265). Nossos resultados mostraram que a deleção do gene zapB provocou a formação de cadeias e aumento do comprimento das células, o que indica que este gene participa na divisão em X. citri. Além disso, a deleção do gene zapA não provocou alterações no fenótipo celular. Tanto a ausência de ZapA quanto de ZapB não alteraram a viabilidade celular, indicando que essas proteínas não são essenciais para a sobrevivência de X. citri. A expressão da fusão GFP-ZapB nas células de ΔzapA mostraram que o recrutamento da proteína ZapB para o septo de divisão em X. citri é dependente de ZapA. Em relação a virulência, a ausência dessas proteínas não prejudicou a capacidade de infecção de plantas hospedeiras. Esse trabalho mostrou que as proteínas ZapA e ZapB em X. citri participam dos processos de divisão celular e fornecem subsídios para um melhor entendimento deste processo neste fitopatógeno.
The bacterium Xanthomonas citri subsp. citri (X. citri) is a phytopathogen that attacks all commercial varieties of citrus, causing a significant impact on the worldwide production of oranges. The disease caused by this bacterium is known as citrus canker, which produces cortical lesions in the fruit, decreasing their productivity and quality. To date, there is no cure for citrus canker, and in order to control the disease, integrated management measures are used to avoid major economic losses. In this context, the study of the biology of the pathogen enables the development of control strategies using antibacterial compounds capable of inhibiting vital processes such as cell division. The machinery responsible for the bacterial division is formed mainly by the FtsZ protein, an ancestral tubulin that forms the Z-ring in the center of dividing cells. The Z-ring recruits several proteins that will act in the division process, as well as others proteins with an accessory function that modulates its assembly. Characterizing the function of these proteins is a way of expanding our targets for the development of new compounds that inhibit cell division and may present extra alternatives for controlling citrus canker. In this work, we characterize two accessory proteins, ZapA and ZapB, encoded by this pathogen. We investigated its functions by evaluating the X. citri mutant phenotype with zapB (XAC_RS17260) and zapA (XAC_RS17265) deletion. Our results showed that the deletion of the zapB gene caused the formation of chains and increased cell length, which indicates that this gene participates in the division in X. citri. In addition, the deletion of the zapA gene did not cause changes in the cell phenotype. Both absence of ZapA and ZapB did not alter cell viability, indicating that these proteins are not essential for the survival of X. citri. The expression of the GFP-ZapB fusion in ΔzapA cells showed that recruitment of the ZapB protein to the X. citri-dividing septum is ZapA-dependent. Regarding virulence, the absence of these proteins did not impair the ability of X. citri to infect host plants. This work showed that the proteins ZapA and ZapB in X. citri participate in the processes of cell division and contributes for a better understanding of this process in this phytopathogen.
The bacterium Xanthomonas citri subsp. citri (X. citri) is a phytopathogen that attacks all commercial varieties of citrus, causing a significant impact on the worldwide production of oranges. The disease caused by this bacterium is known as citrus canker, which produces cortical lesions in the fruit, decreasing their productivity and quality. To date, there is no cure for citrus canker, and in order to control the disease, integrated management measures are used to avoid major economic losses. In this context, the study of the biology of the pathogen enables the development of control strategies using antibacterial compounds capable of inhibiting vital processes such as cell division. The machinery responsible for the bacterial division is formed mainly by the FtsZ protein, an ancestral tubulin that forms the Z-ring in the center of dividing cells. The Z-ring recruits several proteins that will act in the division process, as well as others proteins with an accessory function that modulates its assembly. Characterizing the function of these proteins is a way of expanding our targets for the development of new compounds that inhibit cell division and may present extra alternatives for controlling citrus canker. In this work, we characterize two accessory proteins, ZapA and ZapB, encoded by this pathogen. We investigated its functions by evaluating the X. citri mutant phenotype with zapB (XAC_RS17260) and zapA (XAC_RS17265) deletion. Our results showed that the deletion of the zapB gene caused the formation of chains and increased cell length, which indicates that this gene participates in the division in X. citri. In addition, the deletion of the zapA gene did not cause changes in the cell phenotype. Both absence of ZapA and ZapB did not alter cell viability, indicating that these proteins are not essential for the survival of X. citri. The expression of the GFP-ZapB fusion in ΔzapA cells showed that recruitment of the ZapB protein to the X. citri-dividing septum is ZapA-dependent. Regarding virulence, the absence of these proteins did not impair the ability of X. citri to infect host plants. This work showed that the proteins ZapA and ZapB in X. citri participate in the processes of cell division and contributes for a better understanding of this process in this phytopathogen.
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Palavras-chave
Mutagenese, Cancro (Fitopatologia), Genética microbiana, Mutagenesis, Canker (Phytopathology), Microbial Genetics