A two-step scaffolding model for mitotic chromosome assembly.
Topoisomerase IIalpha (topoIIalpha) and 13S condensin are both required for mitotic chromosome assembly. Here we show that they constitute the two main components of the chromosomal scaffold on histone-depleted chromosomes. The structural stability and chromosomal shape of the scaffolding toward harsh extraction procedures are shown to be mediated by ATP or its nonhydrolyzable analogs, but not ADP. TopoIIalpha and 13S condensin components immunolocalize to a radially restricted, longitudinal scaffolding in native-like chromosomes. Double staining for topoIIalpha and condensin generates a barber pole appearance of the scaffolding, where topoIIalpha- and condensin-enriched "beads" alternate; this structure appears to be generated by two juxtaposed, or coiled, chains. Cell cycle studies establish that 13S condensin appears not to be involved in the assembly of prophase chromatids; they lack this complex but contain a topoIIalpha-defined (-mediated?) scaffolding. Condensin associates only during the pro- to metaphase transition. This two-step assembly process is proposed to generate the barber pole appearance of the native-like scaffolding.
Maeshima
Kazuhiro
K
Department of Biochemistry, NCCR Frontiers in Genetics, University of Geneva, 30, Quai Ernest-Ansermet, CH-1211 Geneva 4, Switzerland.
Laemmli
Ulrich K
UK
D000244:Adenosine Diphosphate / Q000235:genetics / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000235:genetics / Q000378:metabolism; D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000201:enzymology* / Q000648:ultrastructure; D002842:Chromatids / Q000235:genetics / Q000648:ultrastructure; D002875:Chromosomes / Q000201:enzymology* / Q000648:ultrastructure; D004250:DNA Topoisomerases, Type II / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005057:Eukaryotic Cells / Q000201:enzymology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008867:Microtomy; D008938:Mitosis / Q000235:genetics*; D008954:Models, Biological; D015394:Molecular Structure; D046912:Multiprotein Complexes; D011418:Prophase / Q000235:genetics
D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; C108312:condensin complexes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D004250:DNA Topoisomerases, Type II