Skip to main content

Advertisement

SpringerLink
Log in

Visualizing the dynamics of chromosome structure formation coupled with DNA replication

  • Research Article
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

A basic question of cell biology is how DNA folds to chromosome. Numbers of examples have suggested the involvement of DNA replication in chromosome structure formation. To visualize and identify the dynamics of chromosome structure formation and to elucidate the involvement of DNA replication in chromosome construction, Cy3-2′-deoxyuridine-5′-triphosphate direct-labeled active replicating DNA was observed in prematurely condensed chromosomes (PCCs) under a confocal scanning microscope utilized with drug-induced premature chromosome condensation (PCC) technique that facilitates the visualization of interphase chromatin as condensed chromosome form. S-phase PCCs revealed clearly the drastic dynamics of chromosome formation that transits during S-phase from a ‘cloudy nebula’ to numerous numbers of ‘beads on a string’ and finally to ‘striped arrays of banding structured chromosome’ along with the progress of DNA replication. The number, distribution, and shape of replication foci were also measured in individual subphases of S-phase more precisely than reported previously; maximally, ∼1,400 foci of 0.35 μm average radius size were scored at the beginning of the S-phase, and the number reduced to ∼100 at the end of the S-phase. Drug-induced PCC clearly provided the new insight that eukaryote DNA replication is tightly coupled with the chromosome condensation/compaction for the construction of the higher-ordered structure of the eukaryote chromosome.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Asakawa Y, Gotoh E (1997) A method for detecting sister chromatid exchanges using prematurely condensed chromosomes and immunogold–silver staining. Mutagenesis 12:175–177

    Article  PubMed  CAS  Google Scholar 

  • Berezney R, Coffey DS (1975) Nuclear protein matrix: association with newly synthesized DNA. Science 189:291–293

    Article  PubMed  CAS  Google Scholar 

  • Berezney R, Dubey DD, Huberman JA (2000) Heterogeneity of eukaryotic replicons, replicon clusters, and replication foci. Chromosoma 108:471–484

    Article  PubMed  CAS  Google Scholar 

  • Bravo R, Macdonald-Bravo H (1987) Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol 105:1549–1554

    Article  PubMed  CAS  Google Scholar 

  • Buongiorno-Nardelli M, Micheli G, Carri MT, Marilley M (1982) A relationship between replicon size and supercoiled loop domains in the eukaryotic genome. Nature 298:100–102

    Article  PubMed  CAS  Google Scholar 

  • Christensen TW, Tye BK (2003) Drosophila MCM10 interacts with members of the prereplication complex and is required for proper chromosome condensation. Mol Biol Cell 14:2206–2215

    Article  PubMed  CAS  Google Scholar 

  • Cook PR (1995) A chromomeric model for nuclear and chromosome structure. J Cell Sci 108:2927–2935

    PubMed  CAS  Google Scholar 

  • Cook PR (1999) The organization of replication and transcription. Science 284:1790–1795

    Article  PubMed  CAS  Google Scholar 

  • Cremer T, Kurz A, Zirb el R, Dietzel S, Rinke B, Schrock E, Speicher MR, Mathieu U, Jauch A, Emmerrich P, Scherthan H, Ried T, Cremer C, Lichter P (1993) The role of chromosome territories in the functional compartmentalization of the cell nucleus. Cold Spring Harbor Symp Quant Biol 58:777–792

    PubMed  CAS  Google Scholar 

  • Durante M, Furusawa Y, Gotoh E (1998) A simple method for simultaneous interphase–metaphase chromosome analysis in biodosimetry. Int J Radiat Biol 74:457–462

    Article  PubMed  CAS  Google Scholar 

  • Edenberg HJ, Huberman JA (1975) Eukaryotic chromosome replication. Annu Rev Genet 9:245–284

    Article  PubMed  CAS  Google Scholar 

  • El Achkar E, Gerbault-Seureau M, Muleris M, Dutrillaux B, Debatisse M (2005) Premature condensation induces breaks at the interface of early and late replicating chromosome bands bearing common fragile sites. Proc Natl Acad Sci USA 102:18069–18074

    Article  PubMed  CAS  Google Scholar 

  • Essers J, Theil AF, Baldeyron C, van Cappellen WA, Houtsmuller AB, Kanaar R, Vermeulen W (2005) Nuclear dynamics of PCNA in DNA replication and repair. Mol Cell Biol 25:9350–9359

    Article  PubMed  CAS  Google Scholar 

  • Fakan S, Hancock R (1974) Localization of newly-synthesized DNA in a mammalian cell as visualized by high resolution autoradiography. Exp Cell Res 83:95–102

    Article  PubMed  CAS  Google Scholar 

  • Ferreira J, Paolella G, Ramos C, Lamond AI (1997) Spatial organization of large-scale chromatin domains in the nucleus: a magnified view of single chromosome territories. J Cell Biol 139:1597–1610

    Article  PubMed  CAS  Google Scholar 

  • Gerlich D, Ellenberg J (2003) Dynamics of chromosome positioning during the cell cycle. Curr Opin Cell Biol 15:664–671

    Article  PubMed  CAS  Google Scholar 

  • Gerlich D, Beaudouin J, Kalbfuss B, Daigle N, Eils R, Ellenberg J (2003) Global chromosome positions are transmitted through mitosis in mammalian cells. Cell 112:751–764

    Article  PubMed  CAS  Google Scholar 

  • Gollin SM, Wray W, Hanks SK, Hittelman WN, Rao PN (1984) The ultrastructural organization of prematurely condensed chromosomes. J Cell Sci Suppl 1:203–221

    PubMed  CAS  Google Scholar 

  • Gotoh E, Asakawa Y (1996) Detection and evaluation of chromosomal aberrations induced by high doses of gamma-irradiation using immunogold–silver painting of prematurely condensed chromosomes. Int J Radiat Biol 70:517–520

    Article  PubMed  CAS  Google Scholar 

  • Gotoh E, Durante M (2006) Chromosome condensation outside of mitosis: mechanisms and new tools. J Cell Physiol 209:297–304 (and cover page)

    Article  PubMed  CAS  Google Scholar 

  • Gotoh E, Tanno Y (2005) Simple biodosimetry method for cases of high-dose radiation exposure using the ratio of the longest/shortest length of Giemsa-stained drug-induced prematurely condensed chromosomes (PCC). Int J Radiat Biol 81:379–385

    Article  PubMed  CAS  Google Scholar 

  • Gotoh E, Asakawa Y, Kosaka H (1995) Inhibition of protein serine/threonine phophatases directly induces premature chromosome condensation in mammalian somatic cells. Biomed Res 16:63–68

    CAS  Google Scholar 

  • Gotoh E, Kawata T, Durante M (1999) Chromatid break rejoining and exchange aberration formation following gamma-ray exposure: analysis in G2 human fibroblasts by chemically induced premature chromosome condensation. Int J Radiat Biol 75:1129–1135

    Article  PubMed  CAS  Google Scholar 

  • Hadlaczky G, Went M, Ringertz NR (1986) Direct evidence for the non-random localization of mammalian chromosomes in the interphase nucleus. Exp Cell Res 167:1–15

    Article  PubMed  CAS  Google Scholar 

  • Hameister H, Sperling K (1984) Description of a chromosome replication unit in individual prematurely condensed human S-phase chromosomes. Chromosoma 90:389–393

    Article  PubMed  CAS  Google Scholar 

  • Hand R (1978) Eucaryotic DNA: organization of the genome for replication. Cell 15:317–325

    Article  PubMed  CAS  Google Scholar 

  • Hanks SK, Rao PN (1980) Initiation of DNA synthesis in the prematurely condensed chromosomes of G1 cells. J Cell Biol 87:285–291

    Article  PubMed  CAS  Google Scholar 

  • Hearst JE, Kauffman L, McClain WM (1998) A simple mechanism for the avoidance of entanglement during chromosome replication. Trends Genet 14:244–247

    Article  PubMed  CAS  Google Scholar 

  • Hittelman WN (1986) Visualization of chromatin events during DNA excision repair in XP cells: deficiency in localized but not generalized chromatin events. Carcinogenesis 7:1975–1980

    Article  PubMed  CAS  Google Scholar 

  • Hittelman WN, Pollard M (1984) Visualization of chromatin events associated with repair of ultraviolet light-induced damage by premature chromosome condensation. Carcinogenesis 5:1277–1285

    Article  PubMed  CAS  Google Scholar 

  • Hittelman WN, Rao PN (1976) Premature chromosome condensation. Conformational changes of chromatin associated with phytohemagglutinin stimulation of peripheral lymphocytes. Exp Cell Res 100:219–222

    Article  PubMed  CAS  Google Scholar 

  • Hozak P, Hassan AB, Jackson DA, Cook PR (1993) Visualization of replication factories attached to nucleoskeleton. Cell 73:361–373

    Article  PubMed  CAS  Google Scholar 

  • Hozak P, Jackson DA, Cook PR (1996) The role of nuclear structure in DNA replication. In: Blow JJ (ed) Eukaryotic DNA replication: frontiers in molecular biology. Oxford Univ. Press, Oxford, pp 124–142

    Google Scholar 

  • Ito S, Gotoh E, Ozawa S, Yanagi K (2002) Epstein–Barr virus nuclear antigen-1 is highly colocalized with interphase chromatin and its newly replicated regions in particular. J Gen Virol 83:2377–2383

    PubMed  CAS  Google Scholar 

  • Jackson DA, Pombo A (1998) Replicon clusters are stable units of chromosome structure: evidence that nuclear organization contributes to the efficient activation and propagation of S phase in human cells. J Cell Biol 140:1285–1295

    Article  PubMed  CAS  Google Scholar 

  • Johnson RT, Rao PN (1970) Mammalian cell fusion: induction of premature chromosome condensation in interphase nuclei. Nature 226:717–722

    Article  PubMed  CAS  Google Scholar 

  • Johnson RT, Rao PN, Hughes HD (1970) Mammalian cell fusion. 3. A HeLa cell inducer of premature chromosome condensation active in cells from a variety of animal species. J Cell Physiol 76:151–157

    Article  PubMed  CAS  Google Scholar 

  • Johnson RT, Gotoh E, Mullinger AM, Ryan AJ, Shiloh Y, Ziv Y, Squires S (1999) Targeting double-strand breaks to replicating DNA identifies a subpathway of DSB repair that is defective in ataxia–telangiectasia cells. Biochem Biophys Res Commun 261:317–325

    Article  PubMed  CAS  Google Scholar 

  • Kireeva N, Lakonishok M, Kireev I, Hirano T, Belmont AS (2004) Visualization of early chromosome condensation: a hierarchical folding, axial glue model of chromosome structure. J Cell Biol 166:775–785

    Article  PubMed  CAS  Google Scholar 

  • Krause SA, Loupart ML, Vass S, Schoenfelder S, Harrison S, Heck MM (2001) Loss of cell cycle checkpoint control in Drosophila Rfc4 mutants. Mol Cell Biol 21:5156–5168

    Article  PubMed  CAS  Google Scholar 

  • Kuroiwa T (1971) Asynchronous condensation of chromosomes from early prophase to late prophase as revealed by electron microscopic autoradiography. Exp Cell Res 69:97–105

    Article  PubMed  CAS  Google Scholar 

  • Lau YF, Arrighi FE (1981) Studies of mammalian chromosome replication. II. Evidence for the existence of defined chromosome replicating units. Chromosoma 83:721–741

    Article  PubMed  CAS  Google Scholar 

  • Loupart ML, Krause SA, Heck MS (2000) Aberrant replication timing induces defective chromosome condensation in Drosophila ORC2 mutants. Curr Biol 10:1547–1556

    Article  PubMed  CAS  Google Scholar 

  • Ma H, Samarabandu J, Devdhar RS, Acharya R, Cheng PC, Meng C, Berezney R (1998) Spatial and temporal dynamics of DNA replication sites in mammalian cells. J Cell Biol 143:1415–1425

    Article  PubMed  CAS  Google Scholar 

  • Ma H, Siegel AJ, Berezney R (1999) Association of chromosome territories with the nuclear matrix. Disruption of human chromosome territories correlates with the release of a subset of nuclear matrix proteins. J Cell Biol 146:531–542

    Article  PubMed  CAS  Google Scholar 

  • Maiorano D, Moreau J, Mechali M (2000) XCDT1 is required for the assembly of pre-replicative complexes in Xenopus laevis. Nature 404:622–625

    Article  PubMed  CAS  Google Scholar 

  • Manders EM, Stap J, Strackee J, van Driel R, Aten JA (1996) Dynamic behavior of DNA replication domains. Exp Cell Res 226: 328–335

    Article  PubMed  CAS  Google Scholar 

  • Manders EM, Kimura H, Cook PR (1999) Direct imaging of DNA in living cells reveals the dynamics of chromosome formation. J Cell Biol 144:813–821

    Article  PubMed  CAS  Google Scholar 

  • Marheineke K, Hyrien O (2001) Aphidicolin triggers a block to replication origin firing in Xenopus egg extracts. J Biol Chem 276:17092–17100

    Article  PubMed  CAS  Google Scholar 

  • Marsden MP, Laemmli UK (1979) Metaphase chromosome structure: evidence for a radial loop model. Cell 17:849–858

    Article  PubMed  CAS  Google Scholar 

  • McHugh B, Heck MM (2003) Regulation of chromosome condensation and segregation. Curr Opin Genet Dev 13:185–190

    Article  PubMed  CAS  Google Scholar 

  • McNeil PL, Warder E (1987) Glass beads load macromolecules into living cells. J Cell Sci 88:669–678

    PubMed  Google Scholar 

  • Mills AD, Blow JJ, White JG, Amos WB, Wilcock D, Laskey RA (1989) Replication occurs at discrete foci spaced throughout nuclei replicating in vitro. J Cell Sci 94:471–477

    PubMed  Google Scholar 

  • Mullinger AM, Johnson RT (1980) Packing DNA into chromosomes. J Cell Sci 46:61–86

    PubMed  CAS  Google Scholar 

  • Mullinger AM, Johnson RT (1983) Units of chromosome replication and packing. J Cell Sci 64:179–193

    PubMed  CAS  Google Scholar 

  • Nakamura H, Morita T, Sato C (1986) Structural organizations of replicon domains during DNA synthetic phase in the mammalian nucleus. Exp Cell Res 165:291–297

    Article  PubMed  CAS  Google Scholar 

  • Nakayasu H, Berezney R (1989) Mapping replicational sites in the eucaryotic cell nucleus. J Cell Biol 108:1–11

    Article  PubMed  CAS  Google Scholar 

  • Nishitani H, Lygerou Z, Nishimoto T, Nurse P (2000) The Cdt1 protein is required to license DNA for replication in fission yeast. Nature 404:625–628

    Article  PubMed  CAS  Google Scholar 

  • O’Keefe RT, Henderson SC, Spector DL (1992) Dynamic organization of DNA replication in mammalian cell nuclei: spatially and temporally defined replication of chromosome-specific alpha-satellite DNA sequences. J Cell Biol 116:1095–1110

    Article  PubMed  CAS  Google Scholar 

  • Olins AL, Olins DE (1974) Spheroid chromatin units (v bodies). Science 183:330–332

    Article  PubMed  CAS  Google Scholar 

  • Pantelias GE, Maillie HD (1983) A simple method for premature chromosome condensation induction in primary human and rodent cells using polyethylene glycol. Somatic Cell Genet 9:533–547

    Article  PubMed  CAS  Google Scholar 

  • Pardoll DM, Vogelstein B, Coffey DS (1980) A fixed site of DNA replication in eucaryotic cells. Cell 19:527–536

    Article  PubMed  CAS  Google Scholar 

  • Pflumm MF (2002) The role of DNA replication in chromosome condensation. BioEssays 24:411–418

    Article  PubMed  CAS  Google Scholar 

  • Pflumm MF, Botchan MR (2001) Orc mutants arrest in metaphase with abnormally condensed chromosomes. Development 128:1697–1707

    PubMed  CAS  Google Scholar 

  • Prasanth SG, Prasanth KV, Siddiqui K, Spector DL, Stillman B (2004) Human Orc2 localizes to centrosomes, centromeres and heterochromatin during chromosome inheritance. EMBO J 23:2651–2663

    Article  PubMed  CAS  Google Scholar 

  • Rao PN (1977) Premature chromosome condensation and the fine structure of chromosomes. In: Yunis JJ (ed) Molecular structure of human chromosomes. Academic, New York, pp 205–231

    Google Scholar 

  • Rao PN, Wilson B, Puck TT (1977) Premature chromosome condensation and cell cycle analysis. J Cell Physiol 91:131–141

    Article  PubMed  CAS  Google Scholar 

  • Sadoni N, Cardoso MC, Stelzer EH, Leonhardt H, Zink D (2004) Stable chromosomal units determine the spatial and temporal organization of DNA replication. J Cell Sci 117:5353–5365

    Article  PubMed  CAS  Google Scholar 

  • Samaniego R, de la Torre C, Moreno Diaz de la Espina S (2002) Dynamics of replication foci and nuclear matrix during S phase in Allium cepa L. cells. Planta 215:195–204

    Article  PubMed  CAS  Google Scholar 

  • Sarkar A, Eroglu S, Poirier MG, Gupta P, Nemani A, Marko JF (2002) Dynamics of chromosome compaction during mitosis. Exp Cell Res 277:48–56

    Article  PubMed  CAS  Google Scholar 

  • Savage JR, Prasad R, Papworth DG (1984) Subdivision of S-phase and its use for comparative purposes in cultured human cells. J Theor Biol 111:355–367

    Article  PubMed  CAS  Google Scholar 

  • Sperling K, Rao PN (1974a) Mammalian cell fusion. V. Replication behaviour of heterochromatin as observed by premature chromosome condensation. Chromosoma 45:121–131

    Article  PubMed  CAS  Google Scholar 

  • Sperling K, Rao PN (1974b) The phenomenon of premature chromosome condensation: its relevance to basic and applied research. Humangenetik 23:235–258

    Article  PubMed  CAS  Google Scholar 

  • Srebniak MI, Trapp GG, Wawrzkiewicz AK, Kazmierczak W, Wiczkowski AK (2005) The usefulness of calyculin A for cytogenetic prenatal diagnosis. J Histochem Cytochem 53:391–394

    Article  PubMed  CAS  Google Scholar 

  • Swedlow JR, Hirano T (2003) The making of the mitotic chromosome: modern insights into classical questions. Mol Cell 11:557–569

    Article  PubMed  CAS  Google Scholar 

  • Terzoudi GI, Malik SI, Pantelias GE, Margaritis K, Manola K, Makropoulos W (2003) A new cytogenetic approach for the evaluation of mutagenic potential of chemicals that induce cell cycle arrest in the G2 phase. Mutagenesis 18:539–543

    Article  PubMed  CAS  Google Scholar 

  • Terzoudi GI, Manola KN, Pantelias GE, Iliakis G (2005) Checkpoint abrogation in G2 compromises repair of chromosomal breaks in ataxia telangiectasia cells. Cancer Res 65:11292–11296

    Article  PubMed  CAS  Google Scholar 

  • Thoma F, Koller T, Klug A (1979) Involvement of histone H1 in the organization of the nucleosome and of the salt-dependent superstructures of chromatin. J Cell Biol 83:403–427

    Article  PubMed  CAS  Google Scholar 

  • Walter J, Schermelleh L, Cremer M, Tashiro S, Cremer T (2003) Chromosome order in HeLa cells changes during mitosis and early G1, but is stably maintained during subsequent interphase stages. J Cell Biol 160:685–697

    Article  PubMed  CAS  Google Scholar 

  • Woodcock CL, Dimitrov S (2001) Higher-order structure of chromatin and chromosomes. Curr Opin Genet Dev 11:130–135

    Article  PubMed  CAS  Google Scholar 

  • Woodcock CL, Frado LL, Rattner JB (1984) The higher-order structure of chromatin: evidence for a helical ribbon arrangement. J Cell Biol 99:42–52

    Article  PubMed  CAS  Google Scholar 

  • Zink D, Cremer T, Saffrich R, Fischer R, Trendelenburg MF, Ansorge W, Stelzer EH (1998) Structure and dynamics of human interphase chromosome territories in vivo. Hum Genet 102:241–251

    Article  PubMed  CAS  Google Scholar 

  • Zink D, Bornfleth H, Visser A, Cremer C, Cremer T (1999) Organization of early and late replicating DNA in human chromosome territories. Exp Cell Res 247:176–188

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Genetic Resources, National Institute of Infectious Diseases, 1-23-1, Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan

    Eisuke Gotoh

  2. Department of Radiology, Jikei University School of Medicine, 3-25-8, Nishi-Shimbashi, Minato-ku, Tokyo, 116, Japan

    Eisuke Gotoh

Authors
  1. Eisuke Gotoh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Eisuke Gotoh.

Additional information

Communicated by J. Diffley

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gotoh, E. Visualizing the dynamics of chromosome structure formation coupled with DNA replication. Chromosoma 116, 453–462 (2007). https://doi.org/10.1007/s00412-007-0109-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00412-007-0109-5

Keywords

Search

Navigation