We have investigated the sequence specificity of DNA damage induced by mitomycin C reduced with NaBH4, by using 3'- or 5'-end labeled DNA fragments of defined sequence. Mitomycin C reduced with NaBH4 induced heat-labile sites in DNA preferentially at specific sequences. The most preferred trinucleotide sequence for induction of heat-labile sites was GGT, followed by GGG, AGT, GAG, GGC and AGG. Active oxygens such as hydroxyl radical and singlet oxygen, and metal ions were involved in the induction of heat-labile sites. DNA was broken at the 3' side of deoxyguanosines and some of deoxyadenosines by heat-treatment. The produced oligonucleotides contained phosphoryl groups at the 5' termini. The 3' termini seemed not to have simple structures.
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A tetravalent peptide efficiently inhibits the intestinal toxicity of heat-labile enterotoxin by targeting the receptor-binding region of the B-subunit pentamer.
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Department of Molecular Life Sciences, Graduate School of Life and Medical Sciences, Doshisha University, Kyoto, Japan. Electronic address:
Infection by enterotoxigenic Escherichia coli (ETEC) causes severe watery diarrhea and dehydration in humans. Heat-labile enterotoxin (LT) is a major virulence factor produced by ETEC. LT is one of AB-type toxins, such as Shiga toxin (Stx) and cholera toxin (Ctx), and the B-subunit pentamer is responsible for high affinity binding to the LT-receptor, ganglioside GM1, through multivalent interaction.
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Centro Universitario de Estudios Microbiológicos y Parasitológicos, Comisión de Investigaciones Científicas, Universidad Nacional de La Plata, La Plata, Buenos Aires , Argentina.
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