Skip to main content
SpringerLink
Log in

Analysis of DNA replication during S-phase by means of dynamic chromosome banding at high resolution

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

The characteristic patterns of dynamic banding (replication banding) were analysed. Extremely high resolution (850 to 1,250 bands per genome) G- and R-band patterns were obtained after 5-bromo-2′-deoxyuridine (BrdUrd) incorporation either during the early or the late S-phase. We synchronized human lymphocytes with high concentrations of thymidine or BrdUrd as blocking agents, followed by low concentrations of BrdUrd or thymidine respectively as releasing agents, and obtained R- or G-band patterns respectively. The dynamic R-and G-band patterns were complementary for all chromosomes, even for the late-replicating X chromosome. There was no overlapping and every part of each chromosome was positively stained by one of the two banding procedures. The complementarity of the two patterns shows that both high thymidine and high BrdUrd concentrations blocked S-phase progression near the R-band to G-band replication transition in the middle of S-phase. Some bands of the inactive X chromosome replicate before this transition concurrently with R-band replication. The 48 different telomeric regions could be classified into 5 distinct morphotypes based upon the distribution of early and late-replicating DNA in each telomeric region. The dynamic band patterns are particularly useful for the study of the structural and physiological organization of chromosomes at high resolution and should prove invaluable for assessing the replication behavior of rearranged chromosomes.

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

Similar content being viewed by others

References

  • Biemont MC, Laurent C, Couturier J, Dutrillaux B (1978) Chronologic de la réplication des bandes des chromosomes sexuels dans les lymphocytes de sujets normaux et anormaux. Ann Génét 21:133–141

    Google Scholar 

  • Camargo M, Cervenka J (1980) Pattern of chromosomal replication in synchronized lymphocytes. I. Evaluation and application of methotrexate block. Hum Genet 54:47–53

    Google Scholar 

  • Camargo M, Cervenka J (1982) Patterns of DNA replication of human chromosomes. II. Replication map and replication model. Am J Hum Genet 34:757–780

    Google Scholar 

  • Drouin R (1988) La condensation de la chromatine et la réplication de l'ADN dans les chromosomes humains à haute résolution. PhD Thesis, Department of Anatomy, Université de Montréal, Montréal

    Google Scholar 

  • Drouin R, Richer C-L (1985) Analysis of high-resolution R-bands, obtained by heat-denaturation and Giemsa staining, on human prophase chromosomes. Can J Genet Cytol 27:83–91

    Google Scholar 

  • Drouin R, Richer C-L (1989) High-resolution R-banding at the 1250-band level II. Schematic representation and nomenclature of human RBG-banded chromosomes. Genome 32:425–439

    Google Scholar 

  • Drouin R, Lemieux N, Richer C-L (1988a) High-resolution Rbanding at the 1250-band level. I. Technical considerations on cell synchronization and R-banding (RHG and RBG). Cytobios 56:107–125

    Google Scholar 

  • Drouin R, Messier P-E, Richer C-L (1988b) Human chromosome banding specific for electron microscopy. Cytogenet Cell Genet 47:117–120

    Google Scholar 

  • Drouin R, Messier P-E, Richer C-L (1989a) Dynamic G-and Rbanding of human chromosomes for electron microscopy. Chromosoma 98:40–48

    Google Scholar 

  • Drouin R, Messier P-E, Richer C-L (1989b) DNA denaturation for ultrastructural banding and the mechanism underlying the fluorochrome-photolysis-Giemsa technique studied with anti-5-bromodeoxyuridine antibodies. Chromosoma 98:174–180

    Google Scholar 

  • Dutrillaux B, Viegas-Péquignot E (1981) High resolution R- and G-banding on the same preparation. Hum Genet 57:93–95

    Google Scholar 

  • Dutrillaux B, Laurent C, Couturier J, Lejeune J (1973) Coloration des chromosomes humains par l'acridine orange après traitement par le 5-bromodéoxyuridine. CR Séances Acad Sci III 276:3179–3181

    Google Scholar 

  • Dutrillaux B, Couturier J, Richer C-L, Viegas-Péquignot E (1976) Sequence of DNA replication in 277 R- and Q-bands of human chromosomes using a BrdU treatment. Chromosoma 58:51–61

    Google Scholar 

  • Gentry GA, Morse PA, Potter R van (1965) Pyrimidine metabolism in tissue culture cells derived from rat hepatomas. III. Relationship of thymidine to the metabolism of other pyrimidine nucleosides in suspension cultures derived from the Novikoff hepatoma. Cancer Res 25:517–525

    Google Scholar 

  • Goldman MA, Holmquist GP, Gray MC, Caston LA, Nag A (1984) Replication timing of genes and middle repetitive sequences. Science 224:686–692

    Google Scholar 

  • Goto K, Maeda S, Kano Y, Sugiyama T (1978) Factors involved in differential Giemsa-staining of sister chromatids. Chromosoma 66:351–359

    Google Scholar 

  • Goyanes VJ, Mendez J (1981) Karyotyping chromosomes by electron microscopy. Condensation-inhibition of G bands in human and chinese hamster chromosomes by a BrdU-Hoechst 33258 treatment. Cancer Genet Cytogenet 4:45–51

    Google Scholar 

  • Grzeschik K-H, Kim MA, Johannsmann R (1975) Late replicating bands of human chromosomes demonstrated by fluorochrome and Giemsa staining. Humangenetik 29:41–59

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Holmquist GP (1987) Role of replication time in the control of tissue-specific gene expression. Am J Hum Genet 40:151–173

    Google Scholar 

  • Holmquist G (1988) DNA sequences in G-bands and R-bands. In: Adolph KW (ed) Chromosomes and chromatin, vol II. CRC Press, Boca Raton, pp 75–121

    Google Scholar 

  • Holmquist G, Gray M, Porter T, Jordan J (1982) Characterization of Giemsa dark- and light-band DNA. Cell 31:121–129

    Google Scholar 

  • ISCN (1981) An international system for human cytogenetic nomenclature — high resolution banding. Birth defects: original article series, vol XVII, no 5. March of Dimes Birth Defects Foundation, New York. Also Cytogenet Cell Genet 31:1–23

    Google Scholar 

  • Kim MA, Johannsmann R, Grzeschik K-H (1975) Giemsa staining of the sites replicating DNA early in human lymphocyte chromosomes. Cytogenet Cell Genet 15:363–371

    Google Scholar 

  • Latt SA (1973) Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc Natl Acad Sci USA 70:3395–3399

    Google Scholar 

  • Lemieux N, Drouin R, Richer C-L (1990) High-resolution dynamic and morphologic G-bandings (GBG and GTG): a comparative study. Hum Genet 85 (in press)

  • Meer B, Hameister H, Cerrillo M (1981) Early and late replication patterns of increased resolution in human lymphocyte chromosomes. Chromosoma 82:315–319

    Google Scholar 

  • Meuth M, Green H (1974) Induction of a deoxycytidineless state in cultured mammalian cells by bromodeoxyuridine. Cell 2:109–112

    Google Scholar 

  • Painter RB (1982) A replication model for sister chromatid exchange. In: Sandberg AA (ed) Sister chromatid exchange. Progress and topics in cytogenetics, vol 2. Alan R. Riss, New York, pp 115–121

    Google Scholar 

  • Perry P, Wolff S (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251:156–158

    Google Scholar 

  • Reddy KS, Savage JRK, Papworth DG (1988) Replication kinetics of X chromosomes in fibroblasts and lymphocytes. Hum Genet 79:44–48

    Google Scholar 

  • Richer C-L, Drouin R, Murer-Orlando M, Jean P (1983) Highresolution idiogram of Giemsa R-banded human prophase chromosomes. Can J Genet Cytol 25:642–650

    Google Scholar 

  • Rueckert RR, Mueller GC (1960) Studies on unbalanced growth in tissue culture. I. Induction and consequences of thymidine deficiency. Cancer Res 20:1584–1591

    Google Scholar 

  • Savage JRK (1977) Assignment of aberration breakpoints in banded chromosomes. Nature 270:513–514

    Google Scholar 

  • Schempp W, Müller U (1982) High resolution replication patterns of the human Y chromosome. Intra- and interindividual variation. Chromosoma 86:229–237

    Google Scholar 

  • Schempp W, Sigwarth I, Vogel W (1978) Demonstration of replication patterns corresponding to G- and R-type banding of chromosomes after partial synchronization of cell cultures with BrdU or dT surplus. Hum Genet 45:199–202

    Google Scholar 

  • Schmidt M (1980) Two phases of DNA replication in human cells. Chromosoma 76:101–110

    Google Scholar 

  • Schmidt M, Stolzmann WM (1984) Replication variants of the human inactive X chromosome. II. Frequency and replication rate relative to the other chromosomes of the complement. Chromosoma 89:68–75

    Google Scholar 

  • Schmidt M, Stolzmann WM, Baranovskaya LI (1982) Replication variants of the human inactive X chromosome. I. Variability within lymphocytes of single individuals. Chromosoma 85:405–412

    Google Scholar 

  • Schwemmle S, Mehnert K, Vogel W (1989) How does inactivation change timing of replication in the human X chromosome? Hum Genet 83:26–32

    Google Scholar 

  • Viegas-Péquignot E, Dutrillaux B (1978) Une méthode simple pour obtenir des prophases et des prométaphases. Ann Génét 21:122–125

    Google Scholar 

  • Vogel W, Schempp W, Sigwarth I (1978) Comparison of thymidine, fluorodeoxyuridine, hydroxyurea, and methotrexate blocking at the G1/S phase transition of the cell cycle, studied by replication patterns. Hum Genet 45:193–198

    Google Scholar 

  • Vogel W, Boldin S, Reisacher A, Speit G (1985) Characterization of chromosome replication during S-phase with bromodeoxyuridine labelling in chinese hamster ovary and HeLa cells. Chromosoma 92:363–368

    Google Scholar 

  • Vogel W, Autenrieth M, Speit G (1986) Detection of bromodeoxyuridine-incorporation in mammalian chromosomes by a bromodeoxyuridine-antibody. I. Demonstration of replication patterns. Hum Genet 72:129–132

    Google Scholar 

  • Wall WJ, Butler LJ (1989) Classification of Y chromosome polymorphisms by DNA content and C-banding. Chromosoma 97:296–300

    Google Scholar 

  • Willard HF, Latt SA (1976) Analysis of deoxyribonucleic acid replication in human X chromosomes by fluorescence microscopy. Am J Hum Genet 28:213–227

    Google Scholar 

  • Yunis JJ, Sawyer JR, Ball DW (1978) The characterization of highresolution G-banded chromosomes of man. Chromosoma 67:293–307

    Google Scholar 

  • Zakharov AF, Egolina NA (1972) Differential spiralization along mammalian mitotic chromosomes. I. BUdR-revealed differentiation in chinese hamster chromosomes. Chromosoma 38:341–365

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Département d'Anatomie, Faculté de Médecine, Université de Montréal, C.P. 6128, Succ. A, H3C 3J7, Montréal, Québec, Canada

    Régen Drouin, Nicole Lemieux & Claude-Lise Richer

  2. Département de Pathologie et Centre de Recherche Pédiatrique, Hôpital Sainte-Justine, H3T 1C5, Montréal, Québec, Canada

    Nicole Lemieux & Claude-Lise Richer

Authors
  1. Régen Drouin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nicole Lemieux
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Claude-Lise Richer
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Drouin, R., Lemieux, N. & Richer, CL. Analysis of DNA replication during S-phase by means of dynamic chromosome banding at high resolution. Chromosoma 99, 273–280 (1990). https://doi.org/10.1007/BF01731703

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01731703

Keywords

Search

Navigation