Dual roles of the 11S regulatory subcomplex in condensin functions.
Condensin is a multisubunit protein complex that reconfigures DNA structure in an ATP-dependent manner in vitro and plays a central role in mitotic chromosome condensation in Xenopus egg cell-free extracts. The Xenopus 13S condensin complex (13SC) is composed of two subcomplexes: an 8S core subcomplex (8SC) consisting of two structural maintenance of chromosomes (SMC) subunits (XCAP-C and -E) and an 11S regulatory subcomplex (11SR) containing three non-SMC subunits (XCAP-D2, -G, and -H). We report here the biochemical and functional dissection of this chromosome condensation machinery. Although both 8SC and 13SC can bind to DNA in vitro and contain the SMC ATPase subunits, only 13SC is active as a DNA-stimulated ATPase and supports ATP-dependent supercoiling activity. In the cell-free extracts, 13SC is the active form that binds to chromosomes and induces their condensation. Neither 11SR nor 8SC alone is able to bind to chromatin. Our results suggest that the non-SMC subunits have dual roles in the regulation of condensin functions: one is to activate SMC ATPases and the other is to allow the holocomplex to associate with chromatin in a mitosis-specific manner.
Kimura
K
K
Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, NY 11724, USA.
Hirano
T
T
D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002457:Cell Extracts; D002843:Chromatin / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D002875:Chromosomes; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes; D009707:Nucleosomes / Q000378:metabolism; D014981:Xenopus
D002457:Cell Extracts; D002843:Chromatin; D004268:DNA-Binding Proteins; D046912:Multiprotein Complexes; D009707:Nucleosomes; C108312:condensin complexes; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases