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“…The presence of MC-DNA cross-links can be detected by a number of techniques, including standard agarose gel electrophoresis. We exploited the ability of the flavoreductase E 3 (31) (see Materials and Methods) to transfer electrons from NADH to MC in the design of an in vitro assay (23) to measure MRD activity. Various concentrations of MRD were incubated with constant concentrations of all other reaction components (MC, NADH, E 3 , and DNA).…”
Section: Vol 179 1997 Mitomycin-binding Drug Resistance Mechanismmentioning
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rupbmjkragerfmgwileyiopcupepmcmbcthiemesagefrontiersapsiucrarxivemeralduhksmucshluniversity-of-gavle
“…The presence of MC-DNA cross-links can be detected by a number of techniques, including standard agarose gel electrophoresis. We exploited the ability of the flavoreductase E 3 (31) (see Materials and Methods) to transfer electrons from NADH to MC in the design of an in vitro assay (23) to measure MRD activity. Various concentrations of MRD were incubated with constant concentrations of all other reaction components (MC, NADH, E 3 , and DNA).…”
Section: Vol 179 1997 Mitomycin-binding Drug Resistance Mechanismmentioning
“…The resistance gene, mcrA, encodes a flavoenzyme (MCRA) that reoxidizes reductively activated MC (23). In another example involving a DNA damaging agent, bleomycin self-resistance has been determined by drug modification (Bat) and binding (BLMA) proteins in Streptomyces verticillus (38).…”
mentioning
“…The use of multiple resistance mechanisms against a single toxic product is not uncommon (10). Streptomyces lavendulae, for example, protects itself against the deleterious effects of mitomycin via three distinct modes: drug inactivation by a flarin adenine dinucleotide-dependent oxidoreductase (34), drug export (51), and drug sequestration (52). That S. coelicolor employs multiple mechanisms to prevent self-destruction by Act would account for the fact that the actII-ORF2/ORF3 actVA-ORF1 triple mutant (which is defective in exporting Act) is viable (5), as is the soxR mutant.…”
Section: Discussionmentioning
“…The antitumor antibiotic mitomycin C (MC) produced by S. lavendulae is a bioreductively activated alkylating agent that crosslinks DNA at 5'CpG sequences. It has been widely used clinically for antitumor therapy (Johnson et al, 1997). A high molecular weight transglutaminase inhibitor has also been purified from the culture filtrate of S. lavendulae Y-200 (Ikura et al, 2000).…”
Section: Insecticidal and Larvicidal Properties Of Actinomycetesmentioning