title: Quantitative kinetic analysis of nucleolar breakdown and reassembly during mitosis in live human cells.
abstract: One of the great mysteries of the nucleolus surrounds its disappearance during mitosis and subsequent reassembly at late mitosis. Here, the relative dynamics of nucleolar disassembly and reformation were dissected using quantitative 4D microscopy with fluorescent protein-tagged proteins in human stable cell lines. The data provide a novel insight into the fates of the three distinct nucleolar subcompartments and their associated protein machineries in a single dividing cell. Before the onset of nuclear envelope (NE) breakdown, nucleolar disassembly started with the loss of RNA polymerase I subunits from the fibrillar centers. Dissociation of proteins from the other subcompartments occurred with faster kinetics but commenced later, coincident with the process of NE breakdown. The reformation pathway also follows a reproducible and defined temporal sequence but the order of reassembly is shown not to be dictated by the order in which individual nucleolar components reaccumulate within the nucleus after mitosis.
authors.affiliation: Division of Gene Regulation and Expression, School of Life Sciences, Wellcome Trust Biocentre, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Anthony Kar Lun
authors.initials: AK
authors.lastname: Leung
authors.forename: Daniel
authors.initials: D
authors.lastname: Gerlich
authors.forename: Gail
authors.initials: G
authors.lastname: Miller
authors.forename: Carol
authors.initials: C
authors.lastname: Lyon
authors.forename: Yun Wah
authors.initials: YW
authors.lastname: Lam
authors.forename: David
authors.initials: D
authors.lastname: Lleres
authors.forename: Nathalie
authors.initials: N
authors.lastname: Daigle
authors.forename: Joost
authors.initials: J
authors.lastname: Zomerdijk
authors.forename: Jan
authors.initials: J
authors.lastname: Ellenberg
authors.forename: Angus I
authors.initials: AI
authors.lastname: Lamond
mesh_terms: D002466:Cell Nucleolus / Q000378:metabolism*; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D021621:Imaging, Three-Dimensional; D007150:Immunohistochemistry; D007700:Kinetics; D008938:Mitosis; D008954:Models, Biological; D009685:Nuclear Envelope / Q000378:metabolism; D009697:Nucleolus Organizer Region / Q000378:metabolism; D011233:Precipitin Tests; D012318:RNA Polymerase I / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D012318:RNA Polymerase I