Reversible inhibition of mammalian tubulin assembly in vitro and effects in Saccharomyces cerevisiae D61.M by mitomycin C.
Gaulden reported a novel and unexpected mitomycin C (MMC) effect, namely a pronounced retardation of very late prophase and loss of chromosome orientation in neuroblasts of the grasshopper Chortophaga viridifasciate. Because this effect may be due to interactions of MMC with non-DNA targets, MMC was tested for its interaction with porcine brain tubulin assembly in vitro and for the induction of chromosomal malsegregation in the diploid yeast Saccharomyces cerevisiae strain D61.M. A reversible dose-dependent inhibition of tubulin assembly was observed. Since no biological activation system was present in the incubation mixture this inhibition seems to result from an interaction of unactivated MMC with the assembly process. The possible chemical activation of MMC by reduction with 1,4-dithioerythritol (DTE) was investigated by omission of this compound during isolation and polymerization of tubulin. The absence of DTE resulted in a strong reduction of the net tubulin assembly. Also under these conditions MMC led to a dose-dependent inhibition of the assembly, indicating that the effect of MMC on tubulin assembly is independent of a reductive chemical modification. In S. cerevisiae D61.M, MMC did not induce chromosome loss, but induced other genetic events (possibly mutations, deletions or mitotic recombination) as was detected by an increase of the total number and of the frequency of cycloheximide-resistant colonies. This effect could be observed with and without the addition of rat liver S9 as an exogenous activation system.
Albertini
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Pharmaceutical Research/Preclinical Development, F. Hoffmann-La Roche & Co. Ltd., Basel, Switzerland.
Friederich
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Würgler
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FE
D000818:Animals; D001923:Brain Chemistry; D002470:Cell Survival; D002872:Chromosome Deletion; D003513:Cycloheximide / Q000633:toxicity; D004171:Diploidy; D004226:Dithioerythritol / Q000008:administration & dosage; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance; D004789:Enzyme Activation; D008099:Liver / Q000201:enzymology; D016685:Mitomycin; D008937:Mitomycins / Q000008:administration & dosage / Q000633:toxicity*; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000187:drug effects*; D013234:Stem Cells; D013552:Swine; D014404:Tubulin / Q000648:ultrastructure*
D008937:Mitomycins; D014404:Tubulin; D016685:Mitomycin; D004226:Dithioerythritol; D003513:Cycloheximide