The reaction of neocarzinostatin chromphore (1) with 4-hydroxythiophenol under DNA-cleaving conditions was investigated. It was found that a novel cyclization product (5) is formed as a major product together with a minor amount of normal cyclization product 4b. Tritium was incorporated into 4b and 5 from [H-3]-labeled poly(dA-dT), implying that both processes giving 4b and 5 are actually involved in the DNA cleavage reaction.