title: Clathrin is required for the function of the mitotic spindle.
abstract: Clathrin has an established function in the generation of vesicles that transfer membrane and proteins around the cell. The formation of clathrin-coated vesicles occurs continuously in non-dividing cells, but is shut down during mitosis, when clathrin concentrates at the spindle apparatus. Here, we show that clathrin stabilizes fibres of the mitotic spindle to aid congression of chromosomes. Clathrin bound to the spindle directly by the amino-terminal domain of clathrin heavy chain. Depletion of clathrin heavy chain using RNA interference prolonged mitosis; kinetochore fibres were destabilized, leading to defective congression of chromosomes to the metaphase plate and persistent activation of the spindle checkpoint. Normal mitosis was rescued by clathrin triskelia but not the N-terminal domain of clathrin heavy chain, indicating that stabilization of kinetochore fibres was dependent on the unique structure of clathrin. The importance of clathrin for normal mitosis may be relevant to understanding human cancers that involve gene fusions of clathrin heavy chain.
authors.affiliation: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK. sjr51@mrc-lmb.cam.ac.uk
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Royle
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Bright
authors.forename: Leon
authors.initials: L
authors.lastname: Lagnado
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000378:metabolism; D002460:Cell Line; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D020090:Chromosome Segregation; D002966:Clathrin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D033922:Clathrin Heavy Chains / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D008938:Mitosis; D017434:Protein Structure, Tertiary; D051381:Rats; D008941:Spindle Apparatus / Q000502:physiology* / Q000648:ultrastructure
chemical_list: D001324:Autoantigens; C497926:CENPB protein, human; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D002966:Clathrin; D004268:DNA-Binding Proteins; D033922:Clathrin Heavy Chains