Caracterização funcional da ORF XAC0239 de Xanthomonas citri subsp. citri
Carregando...
Data
2018-05-25
Autores
Mardegan, Catarina [UNESP]
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
A Xanthomonas citri subsp. citri (Xac) é a bactéria causadora da doença cancro cítrico. A doença atinge todas as variedades comerciais de citros e, o entendimento da relação fitopatogênica permitirá conhecer formas mais eficientes de controle da bactéria. Com o sequenciamento e análise do genoma e transcriptoma da Xac, foram identificadas ORFs que, provavelmente, estão envolvidas em processos de ataque e colonização da bactéria. Para iniciar a caracterização de uma destas ORFs foram utilizadas, neste estudo, as estratégias de mutação sítio-dirigida de PCR por sobreposição e extensão e modelagem molecular. A ORF XAC0239, é predita como proteína de membrana plasmática, e por homologia, possivelmente, pode fazer parte de uma proteína transportadora de zinco, além disso, há indícios de que é regulada por um fator sigma. A mutação teve efeito negativo sobre a ação de celulases e motilidade swimming e efeito positivo sobre a formação de biofilme e a multiplicação bacteriana in vivo. Aqui é sugerido que na falta desta proteína, houve pouca absorção de zinco, necessário para a formação de biofilme e consequentemente, para a patogenicidade. Além disso, sugestiona-se que, como subterfúgio, a bactéria superexpressou genes para produção e/ou ação de celulases, tentando atacar o hospedeiro de forma alternativa.
Xanthomonas citri subsp. citri (Xac) causes the citrus canker disease. The disease reaches all commercial varieties of citrus and, the understanding of the phytopathogenic relationship allow knowing the ways of control of bacteria. With sequencing of the Xac genome and the realization of a transcriptome, ORFs have been identified that are probably involved in bacterial attack and colonization processes. To initiate the characterization of one of these ORFs, in this study, the strategies of site-directed mutagenesis by overlap extension PCR and molecular modeling were used. The ORF XAC0239 is predicted as plasma membrane protein, and by homology may possibly be part of a zinc transporter protein, moreover, there are indications that it is regulated by a sigma factor. The mutation had a negative effect at the cellulases action and swimming motility and a positive effect on biofilm formation and in vivo multiplication. Here it is suggested that in the absence of this protein, there was little absorption of zinc, necessary for biofilm formation and consequently for pathogenicity. In addition, it is suggested that, as a subterfuge, the bacteria overexpressed genes for production and / or cellulases action, trying to attack the host on alternative way.
Xanthomonas citri subsp. citri (Xac) causes the citrus canker disease. The disease reaches all commercial varieties of citrus and, the understanding of the phytopathogenic relationship allow knowing the ways of control of bacteria. With sequencing of the Xac genome and the realization of a transcriptome, ORFs have been identified that are probably involved in bacterial attack and colonization processes. To initiate the characterization of one of these ORFs, in this study, the strategies of site-directed mutagenesis by overlap extension PCR and molecular modeling were used. The ORF XAC0239 is predicted as plasma membrane protein, and by homology may possibly be part of a zinc transporter protein, moreover, there are indications that it is regulated by a sigma factor. The mutation had a negative effect at the cellulases action and swimming motility and a positive effect on biofilm formation and in vivo multiplication. Here it is suggested that in the absence of this protein, there was little absorption of zinc, necessary for biofilm formation and consequently for pathogenicity. In addition, it is suggested that, as a subterfuge, the bacteria overexpressed genes for production and / or cellulases action, trying to attack the host on alternative way.
Descrição
Palavras-chave
Celulose, Fator sigma, Proteína hipotética, Proteína de membrana, Regulação gênica