A role of topoisomerase II in linking DNA replication to chromosome condensation.
The condensin complex and topoisomerase II (topo II) have different biochemical activities in vitro, and both are required for mitotic chromosome condensation. We have used Xenopus egg extracts to investigate the functional interplay between condensin and topo II in chromosome condensation. When unreplicated chromatin is directly converted into chromosomes with single chromatids, the two proteins must function together, although they are independently targeted to chromosomes. In contrast, the requirement for topo II is temporarily separable from that of condensin when chromosome assembly is induced after DNA replication. This experimental setting allows us to find that, in the absence of condensin, topo II becomes enriched in an axial structure within uncondensed chromatin. Subsequent addition of condensin converts this structure into mitotic chromosomes in an ATP hydrolysis-dependent manner. Strikingly, preventing DNA replication by the addition of geminin or aphidicolin disturbs the formation of topo II-containing axes and alters the binding property of topo II with chromatin. Our results suggest that topo II plays an important role in an early stage of chromosome condensation, and that this function of topo II is tightly coupled with prior DNA replication.
Cuvier
Olivier
O
Cold Spring Harbor Laboratory, 1 Bungtown Rd., Cold Spring Harbor, NY 11724, USA.
Hirano
Tatsuya
T
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology* / Q000648:ultrastructure; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D022004:Chromosome Structures / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000166:cytology / Q000201:enzymology*; D005260:Female; D008297:Male; D008938:Mitosis / Q000235:genetics*; D046912:Multiprotein Complexes; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D009865:Oocytes; D013094:Spermatozoa; D013713:Teniposide / Q000494:pharmacology; D014982:Xenopus laevis
D002843:Chromatin; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; D019384:Nucleic Acid Synthesis Inhibitors; C108312:condensin complexes; D013713:Teniposide; D000251:Adenosine Triphosphatases; D004250:DNA Topoisomerases, Type II