Cytological analysis of chromosome structural defects that result from topoisomerase II dysfunction.
For analyzing chromosome structural defects that result from topoisomerase II (topo II) dysfunction, we have adapted classical cell cycle experiments, classical cytological techniques, and the use of a potent topo II inhibitor (ICRF-193). In this chapter, we describe in detail the protocols used and we discuss the rationale for our choice and for the adaptations applied. We clarify in which cell cycle stages each of the different chromosomal aberrations induced by inhibiting topo II take place: lack of chromosome segregation, undercondensation, lack of sister chromatid resolution, and lack of chromosome individualization. We also put these observations into the context of the two topo II-dependent cell cycle checkpoints.
Giménez-Abián
Juan F
JF
Reproducción Celular y Animal, CIB, CSIC, Madrid, Spain.
Clarke
Duncan J
DJ
D002453:Cell Cycle / Q000502:physiology; D002869:Chromosome Aberrations / Q000139:chemically induced / Q000187:drug effects; D002875:Chromosomes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism*; D054659:Diketopiperazines; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D010879:Piperazines / Q000494:pharmacology; D016622:Silver Staining / Q000379:methods*; D008941:Spindle Apparatus / Q000378:metabolism; D059005:Topoisomerase II Inhibitors
D054659:Diketopiperazines; D004791:Enzyme Inhibitors; D010879:Piperazines; D059005:Topoisomerase II Inhibitors; C070899:4,4'-(1,2-dimethyl-1,2-ethanediyl)bis-2,6-piperazinedione; D004250:DNA Topoisomerases, Type II