Dynamic elastic behavior of alpha-satellite DNA domains visualized in situ in living human cells

We have constructed a fluorescent alpha-satellite DNA-binding protein to explore the motile and mechanical properties of human centromeres. A fusion protein consisting of human CENP-B coupled to the green fluorescent protein (GFP) of A. victoria specifically targets to centromeres when expressed in human cells. Morphometric analysis revealed that the alpha-satellite DNA domain bound by CENPB-GFP becomes elongated in mitosis in a microtubule-dependent fashion. Time lapse confocal microscopy in live mitotic cells revealed apparent elastic deformations of the central domain of the centromere that occurred during metaphase chromosome oscillations. These observations demonstrate that the interior region of the centromere behaves as an elastic element that could play a role in the mechanoregulatory mechanisms recently identified at centromeres. Fluorescent labeling of centromeres revealed that they disperse throughout the nucleus in a nearly isometric expansion during chromosome decondensation in telophase and early G1. During interphase, centromeres were primarily stationary, although motility of individual or small groups of centromeres was occasionally observed at very slow rates of 7-10 microns/h.

• Publication year

  1996

• Venue

  Journal of Cell Biology

• Publication date

  1996-11-01

• Fields of study

  Biology, Medicine, Physics

• Identifiers
  DOI 10.1083/JCB.135.3.545PMID 8909532PMCID 2121065
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Light and electron microscopy of rat kangaroo cells in mitosis

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• Separation of sister chromatids in mitosis requires the Drosophila pimples product, a protein degraded after the metaphase/anaphase transition.

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• Homologous Association of Oppositely Imprinted Chromosomal Domains

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• Kinetochore motility after severing between sister centromeres using laser microsurgery: evidence that kinetochore directional instability and position is regulated by tension.

  1995cited by this paper
• A postprophase topoisomerase II-dependent chromatid core separation step in the formation of metaphase chromosomes

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• Organization of centromeric domains in hepatocyte nuclei: rearrangement associated with de novo activation of the vitellogenin gene family in Xenopus laevis.

  1995cited by this paper
• Islands of complex DNA are widespread in Drosophila centric heterochromatin.

  1995cited by this paper
• Transcription Against an Applied Force

  1995cited by this paper
• Minus-end-directed motion of kinesin–coated microspheres driven by microtubule depolymerization

  1995cited by this paper
• Localization of centromere function in a Drosophila minichromosome.

  1995cited by this paper
• Kinetochore chemistry is sensitive to tension and may link mitotic forces to a cell cycle checkpoint

  1995cited by this paper
• Force generation by microtubule assembly/disassembly in mitosis and related movements.

  1995cited by this paper
• 1. EARLY HISTORY: THE DYNAMIC EQUILIBRIUM MODEL

  1995cited by this paper
• Mitotic forces control a cell-cycle checkpoint

  1995cited by this paper
• The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores

  1995cited by this paper
• CENP-F is a protein of the nuclear matrix that assembles onto kinetochores at late G2 and is rapidly degraded after mitosis

  1995cited by this paper
• A novel cis-acting centromeric DNA element affects S. pombe centromeric chromatin structure at a distance

  1995cited by this paper
• The SMC family: from chromosome condensation to dosage compensation.

  1995cited by this paper
• The Centromere: Hub of Chromosomal Activities

  1995cited by this paper
• Elements of error correction in mitosis: microtubule capture, release, and tension

  1994cited by this paper
• De novo formation of several features of a centromere following introduction of a Y alphoid YAC into mammalian cells.

  1994cited by this paper
• Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle

  1994cited by this paper
• Green fluorescent protein as a marker for gene expression.

  1994cited by this paper
• Structural analysis of alpha-satellite DNA and centromere proteins using extended chromatin and chromosomes.

  1994cited by this paper
• Human CENP-A contains a histone H3 related histone fold domain that is required for targeting to the centromere

  1994cited by this paper
• Tetrasomy 15q: two marker chromosomes with no detectable alpha-satellite DNA.

  1994cited by this paper
• A heterodimeric coiled-coil protein required for mitotic chromosome condensation in vitro.

  1994cited by this paper
• Distribution of CENP-B boxes reflected in CREST centromere antigenic sites on long-range alpha-satellite DNA arrays of human chromosome 21.

  1994cited by this paper
• A novel epigenetic effect can alter centromere function in fission yeast.

  1994cited by this paper
• Anaphase is initiated by proteolysis rather than by the inactivation of maturation-promoting factor.

  1993cited by this paper
• A functional marker centromere with no detectable alpha-satellite, satellite III, or CENP-B protein: activation of a latent centromere?

  1993cited by this paper
• Differential expression of a phosphoepitope at the kinetochores of moving chromosomes

  1993cited by this paper
• Role of chromosome territories in the functional compartmentalization of the cell nucleus.

  1993cited by this paper
• Directional instability of kinetochore motility during chromosome congression and segregation in mitotic newt lung cells: a push-pull mechanism

  1993influential reference
• SMC1: an essential yeast gene encoding a putative head-rod-tail protein is required for nuclear division and defines a new ubiquitous protein family

  1993cited by this paper
• Mitotic block in HeLa cells by vinblastine: ultrastructural changes in kinetochore-microtubule attachment and in centrosomes.

  1993cited by this paper
• Localization of DNA sequences required for human centromere function through an analysis of rearranged Y chromosomes

  1993cited by this paper
• Mammalian chromosome structure.

  1993cited by this paper
• Cell cycle-dependent specific positioning and clustering of centromeres and telomeres in fission yeast

  1993cited by this paper
• CENP-E is a putative kinetochore motor that accumulates just before mitosis

  1992cited by this paper
• Primary structure of the Aequorea victoria green-fluorescent protein.

  1992cited by this paper
• Integration of human α-satellite DNA into simian chromosomes: Centromere protein binding and disruption of normal chromosome segregation

  1992cited by this paper
• Identification of a subdomain of CENP-B that is necessary and sufficient for localization to the human centromere

  1992cited by this paper
• Cell cycle dependent chromosomal movement in pre-mitotic human T-lymphocyte nuclei

  1992cited by this paper
• A human centromere protein, CENP-B, has a DNA binding domain containing four potential alpha helices at the NH2 terminus, which is separable from dimerizing activity

  1992cited by this paper
• Structure of DNA near long tandem arrays of alpha satellite DNA at the centromere of human chromosome 7.

  1992cited by this paper
• Effects of vinblastine, podophyllotoxin and nocodazole on mitotic spindles. Implications for the role of microtubule dynamics in mitosis.

  1992cited by this paper
• Nuclear distribution of centromeres during the cell cycle of human diploid fibroblasts.

  1991cited by this paper
• The chromatin structure of centromeres from fission yeast: differentiation of the central core that correlates with function

  1991cited by this paper
• Structural and mechanical control of mitotic progression.

  1991cited by this paper
• CENP-B is a highly conserved mammalian centromere protein with homology to the helix-loop-helix family of proteins

  1991cited by this paper
• Chromosome topology in mammalian interphase nuclei.

  1991cited by this paper
• Evolution of alpha satellite.

  1991cited by this paper
• Identification of novel centromere/kinetochore-associated proteins using monoclonal antibodies generated against human mitotic chromosome scaffolds

  1991cited by this paper
• Two different microtubule-based motor activities with opposite polarities in kinetochores

  1991cited by this paper
• Modulation of microtubule stability by kinetochores in vitro

  1990cited by this paper
• Determination of three-dimensional imaging properties of a light microscope system. Partial confocal behavior in epifluorescence microscopy.

  1990cited by this paper
• CENP-B: a major human centromere protein located beneath the kinetochore

  1990cited by this paper
• A human centromere antigen (CENP-B) interacts with a short specific sequence in alphoid DNA, a human centromeric satellite

  1989cited by this paper
• Tension, microtubule rearrangements, and the proper distribution of chromosomes in mitosis

  1989cited by this paper
• Long-range organization of tandem arrays of alpha satellite DNA at the centromeres of human chromosomes: high-frequency array-length polymorphism and meiotic stability.

  1989cited by this paper
• A map of the centromere (primary constriction) in vertebrate chromosomes at metaphase.

  1989cited by this paper
• Visualization of centromere proteins CENP-B and CENP-C on a stable dicentric chromosome in cytological spreads

  1989cited by this paper
• Focal points for chromosome condensation and decondensation revealed by three-dimensional in vivo time-lapse microscopy

  1989cited by this paper
• Monoclonal antibodies specific for thiophosphorylated proteins recognize Xenopus MPF.

  1988cited by this paper
• Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries

  1988cited by this paper
• The forces that move chromosomes in mitosis.

  1988cited by this paper
• Structure of the major block of alphoid satellite DNA on the human Y chromosome.

  1987cited by this paper
• Molecular cloning of cDNA for CENP-B, the major human centromere autoantigen

  1987influential reference
• Properties of the kinetochore in vitro. II. Microtubule capture and ATP- dependent translocation

  1985cited by this paper
• Indications of Centromere Movement during Interphase and Differentiation a

  1985cited by this paper
• Different central nervous system cell types display distinct and nonrandom arrangements of satellite DNA sequences.

  1984cited by this paper
• Chromatin conformation of yeast centromeres

  1984cited by this paper
• Measurements of the force produced by the mitotic spindle in anaphase

  1983cited by this paper
• Kinetochore structure, duplication, and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients

  1981cited by this paper
• The attachment of kinetochores to the pro-metaphase spindle in PtK1 cells

  1981cited by this paper
• Cell division in two large pennate diatoms Hantzschia and Nitzschia III. A new proposal for kinetochore function during prometaphase

  1980cited by this paper
• Isolation of a yeast centromere and construction of functional small circular chromosomes

  1980cited by this paper
• Structure and physiology of the mammalian mitotic spindle.

  1975cited by this paper

• Ipsilateral restriction of chromosome movement along a centrosome, and apical-basal axis during the cell cycle

  2025cites this paper
• Prolonged metaphase II arrest weakens Aurora B/C- dependent error correction in mouse oocytes

  2025cites this paper
• Phosphoinositide signaling at the cytoskeleton in the regulation of cell dynamics

  2025cites this paper
• Microtubule-Targeting Agents: Disruption of the Cellular Cytoskeleton as a Backbone of Ovarian Cancer Therapy.

  2024cites this paper
• Ipsilateral restriction of chromosome movement along a centrosome, and apical-basal axis during the cell cycle

  2023influential citation
• Acute irradiation induces a senescence-like chromatin structure in mammalian oocytes

  2023cites this paper
• Exploiting mechanism-informed phenotypic screening for development of next-generation antimitotic phytochemicals

  2021cites this paper
• DNA binding fluorescent proteins as single-molecule probes.

  2020cites this paper
• Genome (in)stability at tandem repeats.

  2020cites this paper
• Loss of sister kinetochore co-orientation and peri-centromeric cohesin protection after meiosis I depends on cleavage of centromeric REC8

  2020cites this paper
• The centromere comes into focus: from CENP-A nucleosomes to kinetochore connections with the spindle

  2020cites this paper
• Polymer perspective of genome mobilization.

  2020cites this paper
• Processive DNA synthesis is associated with localized decompaction of constitutive heterochromatin at the sites of DNA replication and repair

  2019cites this paper
• Molecular Imaging with Reporter Genes: Has Its Promise Been Delivered?

  2019cites this paper
• Novel routes to defined post translational modifications using non-canonical amino acids

  2018cites this paper
• Applications of the CRISPR/Cas system beyond gene editing

  2018cites this paper
• Spo13 prevents premature cohesin cleavage during meiosis

  2018cites this paper
• Microinjection of Antibodies Targeting the Lamin A/C Histone-Binding Site Blocks Mitotic Entry and Reveals Separate Chromatin Interactions with HP1, CenpB and PML

  2017cites this paper
• Organization of nuclear architecture during adipocyte differentiation

  2016cites this paper
• CRISPR/Cas9 in Genome Editing and Beyond.

  2016cites this paper
• Differences in the origins of kinetochore-positive and kinetochore-negative micronuclei: A live cell imaging study.

  2016cites this paper
• Chromosomes at Work: Organization of Chromosome Territories in the Interphase Nucleus

  2016cites this paper
• Proliferation-dependent positioning of individual centromeres in the interphase nucleus of human lymphoblastoid cell lines

  2015cites this paper
• Targeting Microtubules by Natural Agents for Cancer Therapy

  2014cites this paper
• Cell Type Specific Alterations in Interchromosomal Networks across the Cell Cycle

  2014cites this paper
• Dynamique de l'organisation nucléaire des séquences d'ADN répétées centromériques humaines au cours du cycle cellulaire

  2014cites this paper
• Mammalian satellite DNA: a speaking dumb.

  2013cites this paper
• Analysis of a secondary constriction in choromosome 5R of RYE and its neocentromeric activity. Análisis de una constricción secundaria en el cromosoma 5R de centeno y su actividad neocentromérica

  2013cites this paper
• Kif18A and chromokinesins confine centromere movements via microtubule growth suppression and spatial control of kinetochore tension.

  2012cites this paper
• Chromatin mobility is increased at sites of DNA double-strand breaks

  2012cites this paper
• p31comet acts to ensure timely spindle checkpoint silencing subsequent to kinetochore attachment

  2011cites this paper
• Label-Free 3 D Visualization of Cellular and Tissue Structures in Intact Muscle with Second and Third Harmonic Generation Microscopy

  2011cites this paper
• Live-cell visualisation of chromatin for the study of radiation-induced chromosome aberrations

  2011influential citation
• Spatial arrangement of the human genome and its possible functional role

  2011cites this paper
• Tumor imaging with multicolor fluorescent protein expression

  2011cites this paper
• Label-Free 3D Visualization of Cellular and Tissue Structures in Intact Muscle with Second and Third Harmonic Generation Microscopy

  2011cites this paper
• Chromatin higher-order structure and dynamics.

  2010cites this paper
• Kinetochore-microtubule dynamics and attachment stability.

  2010cites this paper
• Chromatin dynamics.

  2010influential citation
• Assays for mitotic chromosome condensation in live yeast and mammalian cells

  2009cites this paper
• The ‘kinetochore maintenance loop’—The mark of regulation?

  2009cites this paper
• The role of APC in mitosis and in chromosome instability.

  2009cites this paper
• 3D fluorescent in situ hybridization using Arabidopsis leaf cryosections and isolated nuclei

  2009cites this paper
• Carbendazim Inhibits Cancer Cell Proliferation by Suppressing Microtubule Dynamics

  2009cites this paper
• Rapid telomere motions in live human cells analyzed by highly time-resolved microscopy

  2008cites this paper
• DNA topoisomerase II is a determinant of the tensile properties of yeast centromeric chromatin and the tension checkpoint.

  2008cites this paper
• Kinetochore structure and spindle assembly checkpoint signaling in the budding yeast, Saccharomyces cerevisiae.

  2008cites this paper
• In vivo real‐time imaging of nuclear‐cytoplasmic dynamics of dormancy, proliferation and death of cancer cells  

  2008cites this paper
• Inhibition of centromere dynamics by eribulin (E7389) during mitotic metaphase

  2008cites this paper
• Quantitative determination of the reduction of phototoxicity and photobleaching by controlled light exposure microscopy

  2008cites this paper
• Midzone activation of aurora B in anaphase produces an intracellular phosphorylation gradient

  2008cites this paper
• Displacement and re-accumulation of centromeric cohesin during transient pre-anaphase centromere splitting

  2007cites this paper
• Centromeric DNA hypomethylation as an epigenetic signature discriminates between germ and somatic cell lineages.

  2007cites this paper
• An epigenetic mark generated by the incorporation of CENP-A into centromeric nucleosomes

  2007cites this paper
• Interphase Chromosome Arrangement in Arabidopsis thaliana Is Similar in Differentiated and Meristematic Tissues and Shows a Transient Mirror Symmetry After Nuclear Division

  2007cites this paper
• Time-lapse and retrospective analysis of DNA methylation in mouse preimplantation embryos by live cell imaging.

  2007cites this paper
• Pax6p46 binds chromosomes in the pericentromeric region and induces a mitosis defect when overexpressed.

  2007cites this paper
• MCAK facilitates chromosome movement by promoting kinetochore microtubule turnover

  2007cites this paper
• How do microtubule-targeted drugs work? An overview.

  2007cites this paper
• Depletion of topoisomerase IIα leads to shortening of the metaphase interkinetochore distance and abnormal persistence of PICH-coated anaphase threads

  2007cites this paper
• Moving chromatin within the interphase nucleus-controlled transitions?

  2007cites this paper
• Proteome and phosphoproteome analysis of chromatin associated proteins in rice (Oryza sativa)

  2007cites this paper
• THE GENOME : Joining , Resolving , and Separating Sister Chromatids During Mitosis and Meiosis

  2006cites this paper
• Suppression of microtubule dynamics by benomyl decreases tension across kinetochore pairs and induces apoptosis in cancer cells

  2006cites this paper
• Nuclear organisation in stem cells

  2006cites this paper
• An Artificially Constructed De Novo Human Chromosome Behaves Almost Identically to Its Natural Counterpart during Metaphase and Anaphase in Living Cells

  2006cites this paper
• Misorientation and reduced stretching of aligned sister kinetochores promote chromosome missegregation in EB1‐ or APC‐depleted cells

  2006influential citation
• Condensin I stabilizes chromosomes mechanically through a dynamic interaction in live cells.

  2006cites this paper
• Chromosome Structures: Visualization

  2006cites this paper
• Regulation of chromosome attachment and dynamics by Saccharomyces cerevisiae kinetochores

  2006cites this paper
• Preservation of large-scale chromatin structure in FISH experiments

  2006cites this paper
• Advances in fluorescent tracking of nucleic acids in living cells.

  2006cites this paper
• Studying and manipulating chromatin motion in mammalian cells

  2006cites this paper
• Analysis of a RanGTP-regulated gradient in mitotic somatic cells

  2006cites this paper
• Chromosome Organization within the Nucleus

  2006cites this paper
• Dissecting cellular activity from single genes to single mRNAs

  2006cites this paper
• Replication of centromeric heterochromatin in mouse fibroblasts takes place in early, middle, and late S phase

  2005cites this paper
• Methyl CpG–binding proteins induce large-scale chromatin reorganization during terminal differentiation

  2005cites this paper
• Chromatin organization in the mammalian nucleus.

  2005cites this paper
• In vivo dynamics and kinetics of pKi-67: transition from a mobile to an immobile form at the onset of anaphase.

  2005cites this paper
• The Condensin I Subunit Barren/CAP-H Is Essential for the Structural Integrity of Centromeric Heterochromatin during Mitosis

  2005cites this paper
• [Centromere elasticity].

  2005cites this paper
• Élasticité du centromère

  2005cites this paper
• Centromere positioning and dynamics in living Arabidopsis plants.

  2005cites this paper
• Konfokale Mikroskopie in der Genomforschung

  2005cites this paper
• The function of nuclear architecture: a genetic approach.

  2004cites this paper
• Spatial arrangement of genes, centromeres and chromosomes in human blood cell nuclei and its changes during the cell cycle, differentiation and after irradiation

  2004cites this paper
• Organisation de la chromatine au sein du noyau interphasique : l’art du rangement fonctionnel

  2004cites this paper
• Cellular Dynamics Visualized in Live Cells in Vitro and in Vivo by Differential Dual-Color Nuclear-Cytoplasmic Fluorescent-Protein Expression

  2004cites this paper
• Hec1 and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites.

  2004cites this paper
• Higher order chromatin architecture in the cell nucleus: on the way from structure to function

  2004cites this paper
• Positioning the genome within the nucleus

  2004cites this paper
• Nuclear organization of centromeric domains is not perturbed by inhibition of histone deacetylases

  2004cites this paper
• Visualizing chromosome structure/organization.

  2004cites this paper
• Microtubules as a target for anticancer drugs

  2004cites this paper
• Mammalian genome organization and its implications for the development of gene therapy vectors Review Article

  2004cites this paper
• The radial positioning of chromatin is not inherited through mitosis but is established de novo in early G1.

  2004cites this paper
• NUCLEAR ARCHITECTURE: A Genetic Approach

  2004cites this paper
• Visualization of Prekinetochore Locus on the Centromeric Region of Highly Extended Chromatin Fibers: Does Kinetochore Autoantigen CENP-C Constitute a Kinetochore Organizing Center?

  2004cites this paper
• Analysis of a spindle pole body mutant reveals a defect in biorientation and illuminates spindle forces.

  2004cites this paper