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The <jats:italic>Escherichia\u00a0coli<\/jats:italic> protein toxin cytotoxic necrotizing factor 1 (<jats:styled-content style=\"fixed-case\">CNF<\/jats:styled-content>1) has recently been reported to influence mitochondrial activity in a mouse model of Rett syndrome and to increase <jats:styled-content style=\"fixed-case\">ATP<\/jats:styled-content> content in the brain tissue of an Alzheimer's disease mouse model. In the present work, the ability of <jats:styled-content style=\"fixed-case\">CNF<\/jats:styled-content>1 to influence mitochondrial activity was investigated in <jats:styled-content style=\"fixed-case\">IEC<\/jats:styled-content>\u20106 normal intestinal crypt cells. In these cells, the toxin was able to induce an increase in cellular <jats:styled-content style=\"fixed-case\">ATP<\/jats:styled-content> content, probably due to an increment of the mitochondrial electron transport chain. In addition, the <jats:styled-content style=\"fixed-case\">CNF<\/jats:styled-content>1\u2010induced Rho <jats:styled-content style=\"fixed-case\">GTP<\/jats:styled-content>ase activity also caused changes in the mitochondrial architecture that mainly consisted in the formation of a complex network of elongated mitochondria. The involvement of the c<jats:styled-content style=\"fixed-case\">AMP<\/jats:styled-content>\u2010dependent protein kinase A signaling pathway was postulated. 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