Maximal chromosome compaction occurs by axial shortening in anaphase and depends on Aurora kinase

Eukaryotic cells must first compact their chromosomes before faithfully segregating them during cell division. Failure to do so can lead to segregation defects with pathological consequences, such as aneuploidy and cancer. Duplicated interphase chromosomes are, therefore, reorganized into tight rods before being separated and directed to the newly forming daughter cells. This vital reorganization of chromatin remains poorly understood. To address the dynamics of mitotic condensation of single chromosomes in intact cells, we developed quantitative assays based on confocal time-lapse microscopy of live mammalian cells stably expressing fluorescently tagged core histones. Surprisingly, maximal compaction was not reached in metaphase, but in late anaphase, after sister chromatid segregation. We show that anaphase compaction proceeds by a mechanism of axial shortening of the chromatid arms from telomere to centromere. Chromatid axial shortening was not affected in condensin-depleted cells, but depended instead on dynamic microtubules and Aurora kinase. Acute perturbation of this compaction resulted in failure to rescue segregation defects and in multilobed daughter nuclei, suggesting functions in chromosome segregation and nuclear architecture.

[1]  J. Swedlow,et al.  Focal points for chromosome condensation and decondensation revealed by three-dimensional in vivo time-lapse microscopy , 1989, Nature.

[2]  J. Swedlow,et al.  Multiple chromosomal populations of topoisomerase II detected in vivo by time-lapse, three-dimensional wide-field microscopy , 1993, Cell.

[3]  G. Borisy,et al.  Kinetochore microtubule dynamics and the metaphase-anaphase transition , 1995, The Journal of cell biology.

[4]  H. Bellen,et al.  Chromatid Segregation at Anaphase Requires the barren Product, a Novel Chromosome-Associated Protein That Interacts with Topoisomerase II , 1996, Cell.

[5]  J. Ellenberg,et al.  Four-dimensional imaging and quantitative reconstruction to analyse complex spatiotemporal processes in live cells , 2001, Nature Cell Biology.

[6]  Hiroshi Kimura,et al.  Kinetics of Core Histones in Living Human Cells , 2001, The Journal of cell biology.

[7]  D. Compton,et al.  The chromokinesin Kid is necessary for chromosome arm orientation and oscillation, but not congression, on mitotic spindles , 2001, The Journal of cell biology.

[8]  P. Jallepalli,et al.  Chromosome segregation and cancer: cutting through the mystery , 2001, Nature Reviews Cancer.

[9]  Roland Eils,et al.  Nuclear Envelope Breakdown Proceeds by Microtubule-Induced Tearing of the Lamina , 2002, Cell.

[10]  Barbara J Meyer,et al.  C. elegans condensin promotes mitotic chromosome architecture, centromere organization, and sister chromatid segregation during mitosis and meiosis. , 2002, Genes & development.

[11]  Kim Nasmyth,et al.  Segregating Sister Genomes: The Molecular Biology of Chromosome Separation , 2002, Science.

[12]  D. Cimini,et al.  Merotelic kinetochore orientation versus chromosome mono-orientation in the origin of lagging chromosomes in human primary cells. , 2002, Journal of cell science.

[13]  J. Ellenberg,et al.  4D imaging to assay complex dynamics in live specimens. , 2003, Nature cell biology.

[14]  D. Spector,et al.  The dynamics of chromosome organization and gene regulation. , 2003, Annual review of biochemistry.

[15]  J. Peters,et al.  The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint , 2003, The Journal of cell biology.

[16]  J. Swedlow,et al.  The making of the mitotic chromosome: modern insights into classical questions. , 2003, Molecular cell.

[17]  Roland Eils,et al.  Global Chromosome Positions Are Transmitted through Mitosis in Mammalian Cells , 2003, Cell.

[18]  D. Spector,et al.  Sequential entry of components of the gene expression machinery into daughter nuclei. , 2003, Molecular biology of the cell.

[19]  Stephen S. Taylor,et al.  Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores , 2003, The Journal of cell biology.

[20]  G. J. Brakenhoff,et al.  Four-dimensional imaging of chromatin dynamics during the assembly of the interphase nucleus , 2004, Chromosome Research.

[21]  D. Koshland,et al.  In vivo requirements for rDNA chromosome condensation reveal two cell-cycle-regulated pathways for mitotic chromosome folding. , 2004, Genes & development.

[22]  Daniel C. Ducat,et al.  Aurora kinases in spindle assembly and chromosome segregation. , 2004, Experimental cell research.

[23]  J. Ellenberg,et al.  Distinct functions of condensin I and II in mitotic chromosome assembly , 2004, Journal of Cell Science.

[24]  J. Lippincott-Schwartz,et al.  The fusome mediates intercellular endoplasmic reticulum connectivity in Drosophila ovarian cysts. , 2004, Molecular biology of the cell.

[25]  F. Stegmeier,et al.  Cdc14 and Condensin Control the Dissolution of Cohesin-Independent Chromosome Linkages at Repeated DNA , 2004, Cell.

[26]  F. Uhlmann,et al.  Cdc14 Phosphatase Induces rDNA Condensation and Resolves Cohesin-Independent Cohesion during Budding Yeast Anaphase , 2004, Cell.

[27]  J. Ellenberg,et al.  Automatic real‐time three‐dimensional cell tracking by fluorescence microscopy , 2004, Journal of microscopy.

[28]  David A. Agard,et al.  Large-scale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro , 1989, Chromosoma.

[29]  A. Jarmuz,et al.  Spindle-independent condensation-mediated segregation of yeast ribosomal DNA in late anaphase , 2005, The Journal of cell biology.

[30]  D. Compton,et al.  Efficient Mitosis in Human Cells Lacking Poleward Microtubule Flux , 2005, Current Biology.

[31]  Lin Tang,et al.  Roscovitine Targets, Protein Kinases and Pyridoxal Kinase*[boxs] , 2005, Journal of Biological Chemistry.

[32]  Thomas Cremer,et al.  Methyl CpG–binding proteins induce large-scale chromatin reorganization during terminal differentiation , 2005, The Journal of cell biology.

[33]  Yves Barral,et al.  The NoCut Pathway Links Completion of Cytokinesis to Spindle Midzone Function to Prevent Chromosome Breakage , 2006, Cell.

[34]  H. Erfle,et al.  High-throughput RNAi screening by time-lapse imaging of live human cells , 2006, Nature Methods.

[35]  Jan Ellenberg,et al.  Dissecting the contribution of diffusion and interactions to the mobility of nuclear proteins. , 2006, Biophysical journal.

[36]  J. Ellenberg,et al.  Condensin I Stabilizes Chromosomes Mechanically through a Dynamic Interaction in Live Cells , 2006, Current Biology.

[37]  A. Lamond,et al.  Condensin and Repo-Man–PP1 co-operate in the regulation of chromosome architecture during mitosis , 2006, Nature Cell Biology.

[38]  J. Ellenberg,et al.  Measuring structural dynamics of chromosomes in living cells by fluorescence microscopy. , 2007, Methods.

[39]  N. Amariglio,et al.  Gene silencing at the nuclear periphery , 2007, The FEBS journal.