title: Cell cycle-regulated recognition of the destruction box of cyclin B by the APC/C in Xenopus egg extracts.
abstract: Substrates for mitotic proteolysis such as cyclin B have a 9 residue destruction motif, the destruction box (D-box). To identify the receptor that specifically binds the D-box, we used affinity chromatography with immobilized D-box matrices. We find that the APC/C from Xenopus egg extracts binds to the D-box of cyclin B, whereas Fizzy (Cdc20) does not. Mutations in the D-box abolished this interaction. We show that this binding is regulated in the cell cycle, such that the APC/C from egg extracts in interphase does not bind to the D-box matrix. Our results suggest that the APC/C forms a stable interaction with the D-box of its substrates in a cell cycle-dependent manner.
authors.affiliation: Cancer Research UK, Clare Hall Laboratories, South Mimms, Herts EN6 3LD, United Kingdom.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamano
authors.forename: Julian
authors.initials: J
authors.lastname: Gannon
authors.forename: Hiro
authors.initials: H
authors.lastname: Mahbubani
authors.forename: Tim
authors.initials: T
authors.lastname: Hunt
mesh_terms: D000818:Animals; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002457:Cell Extracts; D002846:Chromatography, Affinity; D019926:Cyclin B / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D009865:Oocytes; D011494:Protein Kinases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance; D014981:Xenopus
chemical_list: D018797:Cell Cycle Proteins; D002457:Cell Extracts; D019926:Cyclin B; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D011494:Protein Kinases