Skip to main content
SpringerLink
Log in

Identification of the Balbiani ring 2 chromomere and determination of the content and compaction of its DNA

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

The giant puff Balbiani ring 2 (BR 2) in the salivary glands of Chironomus tentans is known to contain transcriptionally active 75S RNA genes. The corresponding chromosomal superstructure, the BR 2 chromomere, has been identified and characterized in the present study. It was demonstrated by cytological methods that BR 2 originates from the broad band IV-3B10, since all the bands except the 3B10 band could be detected as intact bands in the vicinity of BR 2. In situ hybridization of BR 2 RNA to squashed Malpighian tubule chromosomes lacking BR 2, showed that all or almost all the DNA sequences complementary to BR 2 RNA are located in the 3B10 band. The DNA amount in the 3B10 band, designated the BR 2 band, was measured in relation to the DNA content of the whole chromosome set by both direct and indirect microspectrophotometry of Feulgen-stained Malpighian tubule chromosomes. From a determination of the haploid DNA content in sperm cells (0.25 pg), the amount of DNA in a BR 2 chromomere could then be calculated to be 5.1×10−4 pg DNA or 470 kb DNA. A minimum value of the DNA compaction within the BR 2 chromomere was estimated to 380 from the B-form length of BR 2 DNA (160 μm) and the thickness of the BR 2 band (0.42 μm). Since there are only between 1 and 4 75S RNA genes, 37 kb in size, in a BR 2 chromomere, most of the BR 2 DNA must consist of DNA not coding for 75S RNA. The nature of this DNA is discussed in relation to the high average AT-content of Chironomus DNA. The organization of the chromosome fiber in the BR 2 chromomere is considered in relation to the gene activation process in the salivary glands, i.e. the formation of a 75S RNA transcription loop from a portion of the tightly packed chromosome fiber in the BR 2 chromomere.

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

  • Alonso, C., Berendes, H.D.: The location of 5S (ribosomal) RNA genes in genes in Drosophila hydei. Chromosoma (Berl.) 51, 347–356 (1975)

    Google Scholar 

  • Ananiev, E.V., Barsky, V.E.: Localization of RNA synthesis sites in the 1B-3C region of the Drosophila melanogaster X chromosome. Chromosoma (Berl.) 65, 359–371 (1978)

    Google Scholar 

  • Artavanis-Tsakonas, S., Schedl, P., Tschudi, C., Pirrotta, V., Steward, R., Gehring, W.J.: The 5S genes of Drosophila melanogaster. Cell 12, 1057–1067 (1977)

    Google Scholar 

  • Ashburner, M.: Function and structure of polytene chromosomes during insect development. In: Advances in insect physiology. (J.W.L. Beament, J.E. Treherne and V.B. Wigglesworth, eds.) 7, 1–95. London: Academic Press 1970

    Google Scholar 

  • Beermann, W.: Chromomerenkonstanz und spezifische Modifikation der Chromosomenstruktur in der Entwicklung und Organdifferenzierung von Chironomus tentans. Chromosoma (Berl.) 5, 139–198 (1952)

    Google Scholar 

  • Beermann, W.: Chromomeres and genes. In: Developmental Studies on Giant Chromosomes. (W. Beermann, ed.) pp. 1–33. Berlin: Springer 1972

    Google Scholar 

  • Beermann, W.: Directed changes in the pattern of Balbiani ring puffing in Chironomus: Effects of sugar treatment. Chromosoma (Berl.) 41, 297–326 (1973)

    Google Scholar 

  • Beermann, W., Bahr, G.F.: The submicroscopic structure of the Balbiani-ring. Exp. Cell Res. 6, 195–201 (1954)

    Google Scholar 

  • Benyajati, C., Worcel, A.: Isolation, characterization, and structure of the folded interphase genome of Drosophila melanogaster. Cell 9, 393–407 (1976)

    Google Scholar 

  • Berendes, H.D.: Polytene chromosome structure at the submicroscopic level. I. A map of region X, 1-4E of Drosophila melanogaster. Chromosoma (Berl.) 29, 118–130 (1970)

    Google Scholar 

  • Berendes, H.D.: The control of puffing in Drosophila hydei. In: Results and Problems in Cell Differentiation. (W. Beermann, ed.), pp. 181–207. Berlin: Springer 1972

    Google Scholar 

  • Bolund, L., Ringertz, N.R., Harris, H.: Changes in the cytochemical properties of erythrocyte nuclei reactivated by cell fusion. J. Cell Sci. 4, 71–87 (1969)

    Google Scholar 

  • Bonner, J.J., Pardue, M.L.: Ecdysone-stimulated RNA synthesis in imaginai discs of Drosophila melanogaster. Assay by in situ hybridization. Chromosoma (Berl.) 58, 87–99 (1976)

    Google Scholar 

  • Bridges, C.B.: Salivary chromosome maps. With a key to the banding of the chromosomes of Drosophila melanogaster. J. Heredity 26, 60–64 (1935)

    Google Scholar 

  • Callan, H.G., Lloyd, L.: Lampbrush chromosomes of crested newts, Triturus cristatus (Laurenti). Phil. Trans. roy. Soc. (Lond.) B. 243, 135–219 (1960).

    Google Scholar 

  • Cannon, G.B.: Culture of insect salivary glands in a chemically defined medium. Science 146, 1063 (1964)

    Google Scholar 

  • Case, S.T., Daneholt, B.: Cellular and molecular aspects of genetic expression in Chironomus salivary glands. In: Int. Rev. Biochem. (I. Paul, ed.) pp. 45–77. Baltimore: University Park Press 1977

    Google Scholar 

  • Case, S.T., Daneholt, B.: The size of the transcription unit in Balbiani ring 2 or Chironomus tentans as derived from analysis of the primary transcript and 75S RNA. J. molec. Biol. 124, 223–241 (1978)

    Google Scholar 

  • Caspersson, T., Lomakka, G.: Microscale spectroscopy. In: Instrumentation in Biochemistry (T.W. Goodwin, ed.), pp. 25–40. London: Academic Press 1966

    Google Scholar 

  • Daneholt, B.: Transcription in polytene chromosomes. Cell 4, 1–9 (1975)

    Google Scholar 

  • Daneholt, B., Edström, J.-E.: The content of deoxyribonucleic acid in individual polytene chromosomes of Chironomus tentans. Cytogenetics 6, 350–356 (1967)

    Google Scholar 

  • Darnell, I.E.: Ribonucleic acids from animal cells. Bacteriol. Rev. 32, 262–290 (1968)

    Google Scholar 

  • Derksen, J.: Cytological analysis of Drosophila polytene chromosomes. In: Methods in Cell Biology. (G. Stein, J. Stein and L.J. Kleinsmith eds.) 17, 133–140. New York: Academic Press 1978

    Google Scholar 

  • Edström, J.-E., Beermann, W.: The base composition of nucleic acids in chromosomes, puffs, nucleoli, and cytoplasm in Chironomus salivary gland cells. J. Cell Biol. 14, 371–379 (1962)

    Google Scholar 

  • Egyhazi, E.: Inhibition of Balbiani ring RNA synthesis at the initiation level. Proc. nat. Acad. Sci.(Wash.) 73, 947–950 (1975)

    Google Scholar 

  • Finch, J.T., Klug, A.: Solenoidal model for superstructure in chromatin. Proc. nat. Acad. Sci.(Wash.) 73, 1897–1901 (1976)

    Google Scholar 

  • Franke, W.W., Scheer, U., Trendelenburg, M., Zentgraf, H., Spring, H.: Morphology of transcriptionally active chromatin. Cold Spr. Harb. Symp. 42, 755–772 (1978)

    Google Scholar 

  • Fujita, S., Takamoto, K.: Synthesis of messenger RNA on the polytene chromosomes of dipteran salivary glands. Nature (Lond.) 200, 494–495 (1963)

    Google Scholar 

  • Gall, J.G., Callan, H.G.: H3uridine incorporation in lampbrush chromosomes. Proc. nat. Acad. Sci. (Wash.) 48, 562–570 (1962)

    Google Scholar 

  • Gall, J.G., Cohen, E.H., Polan, M.L.: Repetitive DNA sequences in Drosophila. Chromosoma (Berl.) 33, 319–344 (1971)

    Google Scholar 

  • Gilbert, W.: Why genes in pieces? Nature (Lond.) 271, 501 (1978)

    Google Scholar 

  • Gledhill, B.L., Gledhill, M.P., Rigler Jr., R., Ringertz, N.R.: Changes in deoxyribonucleoprotein during spermiogenesis in the bull. Exp. Cell Res. 41, 652–665 (1966)

    Google Scholar 

  • Hollenberg, C.P.: Proportionate representation of rDNA and Balbiani ring DNA in polytene chromosomes of Chironomus tentans. Chromosoma (Berl.) 57, 185–197 (1976)

    Google Scholar 

  • Igó-Kemenes, T., Zachau, H.G.: Domains in chromatin structure. Cold Spr. Harb. Symp. 42, 109–118 (1978)

    Google Scholar 

  • Jamrich, M., Greenleaf, A.L., Bautz, E.K.F.: Localization of RNA polymerase in polytene chromosomes of Drosophila melanogaster. Proc. nat. Acad. Sci. (Wash.) 74, 2079–2083 (1977)

    Google Scholar 

  • Keyl, H.-G.: Duplikationen von Untereinheiten der Chromosomalen DNS während der Evolution von Chironomus thummi. Chromosoma (Berl.) 17, 139–180 (1965)

    Google Scholar 

  • Keyl, H.-G.: Lampbrush chromosomes in spermatocytes of Chironomus. Chromosoma (Berl.) 51, 75–91 (1975)

    Google Scholar 

  • Laemmli, U.K., Cheng, S.M., Adolph, K.W., Paulson, J.R., Brown, J.A., Baumbach, W.R.: Metaphase chromosome structure: The role of nonhistone proteins. Cold Spr. Harb. Symp. 42, 351–360 (1978)

    Google Scholar 

  • Lamb, M.M., Daneholt, B.: Characterization of active transcription units in Balbiani rings of Chironomus tentans. Cell 17, 835–848 (1979)

    Google Scholar 

  • Lambert, B.: Repeated nucleotide sequences in polytene chromosomes. Thesis, Karolinska Institutet, Stockholm 1972

    Google Scholar 

  • Lambert, B.: The total chromosomal distribution of Balbiani ring DNA in Chironomus tentans. Chromosoma (Berl.) 50, 193–200 (1975)

    Google Scholar 

  • Lambert, B., Daneholt, B.: Microanalysis of RNA from defined cellular components. In: Methods in Cell Biology X (D.M. Prescott, ed.), 17–47. New York: Academic Press 1975

    Google Scholar 

  • Lambert, B., Wieslander, L., Daneholt, B., Egyhazi, E., Ringborg, U.: In situ demonstration of DNA hybridizing with chromosomal and nuclear sap RNA in Chironomus tentans. J. Cell Biol. 53, 407–418 (1972)

    Google Scholar 

  • Lawn, R.M., Heumann, J.M., Herrick, G., Prescott, D.M.: The gene-size DNA molecules in Oxytrichia. Cold Spr. Harb. Symp. 42, 483–492 (1978)

    Google Scholar 

  • Lefevre Jr., G.: The relationship between genes and polytene chromosome bands. Ann. Rev. Genet. 8, 51–62 (1974)

    Google Scholar 

  • Lifton, R.P., Goldberg, M.L., Karp, R.W., Hogness, D. S.: The organization of the histone genes in Drosophila melanogaster: Functional and evolutionary implications. Cold Spr. Harb. Symp. 42, 1047–1051 (1978)

    Google Scholar 

  • Marsden, M.P.F., Laemmli, U.K.: Metaphase chromosome structure: Evidence for a radial loop model. Cell 17, 849–858 (1979)

    Google Scholar 

  • Mathis, D., Oudet, P., Chambon, P.: Structure of transcribing chromatin. Progr. Nucleic Acid. Res. (in press, 1980)

  • Nelson, L.G., Derksen, J., Lamb, M. M., Wieslander, L., Daneholt, B.: Suppression of transcription in Balbiani ring 2 and the effect on chromosome structure. In: FEBS 11th Meeting, Copenhagen. (B.F.C. Clark, H. Klenow, J. Zeuthen, eds.) 43, 279–286. Oxford: Pergamon Press 1978

    Google Scholar 

  • Pardue, M. L.: Repeated DNA sequences in the chromosomes of higher organisms. Genetics 79, 159–170 (1975)

    Google Scholar 

  • Pelling, C.: Ribonukleinsäure-Synthese der Riesenchromosomen. Autoradiographische Untersuchungen an Chironomus tentans. Chromosoma (Berl.) 15, 71–122 (1964)

    Google Scholar 

  • Pelling, C.: A replicative and synthetic chromosomal unit — the modern concept of the chromosome. Proc. roy. Soc. Lond. B 164, 279–289 (1966)

    Google Scholar 

  • Ploeg, M. van der, Duijn, P. van, Ploem, J.S.: High-resolution scanning densitometry of photographic negatives of human metaphase chromosomes. I. Instrumentation. Histochemistry 42, 9–29 (1974)

    Google Scholar 

  • Ploeg, M. van der, Broek, K. van den, Smeulders, A.W.M., Vossepoel, A.M., Duijn, P. van: HIDACSYS: Computer programs for interactive scanning cytophotometry. Histochemistry 54, 273–288 (1977)

    Google Scholar 

  • Prescott, D.M., Murti, K. G.: Chromosome structure in ciliated protozoans. Cold Spr. Harb. Symp. 38, 609–618 (1974)

    Google Scholar 

  • Rasch, E.M.: DNA cytophotometry of salivary gland nuclei and other tissue systems in dipteran larvae. In: Introduction to Quantitative Cytochemistry II. (G.L. Wied, G.F. Bahr eds.), pp. 357–397. New York: Academic Press 1970

    Google Scholar 

  • Rasch, E.M., Barr, H.J., Rasch, R.W.: The DNA content of sperm of Drosophila melanogaster. Chromosoma (Berl.) 33, 1–18 (1971)

    Google Scholar 

  • Rudkin, G. T.: Microspectrophotometry of chromosomes. In: Introduction to quantitative cytochemistry. (G.L. Wied, ed.), pp. 387–407. New York: Academic Press 1966

    Google Scholar 

  • Saura, A.O., Sorsa, V.: Electron microscopic analysis of the banding pattern in the salivary gland chromosomes of Drosophila melanogaster. Divisions 30 and 31 of 2L. Hereditas (Lund) 90, 257–267 (1979)

    Google Scholar 

  • Sedat, J., Manuelidis, L.: A direct approach to the structure of eukaryotic chromosomes. Cold Spr. Harb. Symp. 42, 331–350 (1978)

    Google Scholar 

  • Semeshin, V.F., Zhimulev, I.F., Belyaeva, E.S.: Electron microscope autoradiographic study on transcriptional activity of Drosophila melanogaster polytene chromosomes. Chromosoma (Berl.) 73, 163–177 (1979)

    Google Scholar 

  • Serfling, E., Maximovsky, L.F., Wobus, U.: Synthesis and processing of ribosomal ribonucleic acid in salivary gland cells of Chironomus thummi. Europ. J. Biochem. 45, 277–289 (1974)

    Google Scholar 

  • Sorsa, V.: Electron microscopic localization and ultrastructure of certain gene loci in salivary gland chromosomes of Drosophila melanogaster. In: Alfred Benzon Symp. 13. (J. Engberg, H. Klenow, and V. Leick, eds.), pp. 55–71. Copenhagen: Munksgaard 1979

    Google Scholar 

  • Sorsa, V., Green, M. M., Beermann, W.: Cytogenetic fine structure and chromosomal localization of the white gene in Drosophila melanogaster. Nature (Lond.) New Biol. 245, 34–37 (1973)

    Google Scholar 

  • Sorsa, V., Pusa, K., Virrankoski, V., Sorsa, M.: Electron microscopy of an induced “lampbrush stage” of the polytene chromosome. Exp. Cell Res. 60, 466–469 (1970b)

    Google Scholar 

  • Sorsa, M., Sorsa, V.: Electron microscopic studies on band regions in Drosophila salivary chromosomes. Ann. Acad. Sci. Fenn. (Ser. A). IV, 127, 1–8 (1968)

    Google Scholar 

  • Sorsa, V., Sorsa, M., Virrankoski, V., Pusa, K.: An electron microscopic study on alkali-treated salivary gland chromosomes of Drosophila. Ann. Acad. Sci. Fenn. (Ser. A) IV, 166, 1–10 (1970a)

    Google Scholar 

  • Stevens, B.J., Swift, H.: RNA transport from nucleus to cytoplasm in Chironomus salivary glands. J. Cell Biol. 31, 55–78 (1966)

    Google Scholar 

  • Trapp, L.: Instrumentation for recording microspectrophotemetry. In: Introduction to Quantitative Cytochemistry. (G.L. Wied, ed.), pp. 427–435. New York: Academic Press 1966

    Google Scholar 

  • Wieslander, L.: The presence of 5S RNA genes in two consecutive chromosome bands in Chironomus tentans. Molec. Biol. Reports 2, 189–194 (1975)

    Google Scholar 

  • Wieslander, L.: Number and structure of Balbiani ring 75S RNA transcription units in Chironomus tentans. J. molec. Biol. 134, 347–367 (1979)

    Google Scholar 

  • Wobus, U.: Molecular characterization of an insect genome: Chironomus thummi. Europ. J. Biochem. 59, 287–293 (1975)

    Google Scholar 

  • Wolstenholme, D.R., Dawid, I.B., Ristow, H.: An electron microscope study of DNA molecules from Chironomus tentans and Chironomus thummi. Genetics 60, 759–770 (1968)

    Google Scholar 

  • Young, M.W., Judd, B.H.: Nonessential sequences, genes, and the polytene chromosome bands of Drosophila melanogaster. Genetics 88, 723–742 (1978)

    Google Scholar 

  • Zhimulev, I.F., Belyaeva, E.S.: 3H-uridine labelling patterns in the Drosophila melanogaster salivary gland chromosomes X, 2R, and 3L. Chromosoma (Berl.) 49, 219–231 (1975)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Histochemistry and Cytochemistry, University of Leiden, Leiden, The Netherlands

    M. van der Ploeg

  2. Department of Histology and Department of Medical Cell Genetics, Karolinska Institutet, S-104 01, Stockholm, Sweden

    J. Derksen, L. Wieslander & B. Daneholt

Authors
  1. J. Derksen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L. Wieslander
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. van der Ploeg
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B. Daneholt
    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

Derksen, J., Wieslander, L., van der Ploeg, M. et al. Identification of the Balbiani ring 2 chromomere and determination of the content and compaction of its DNA. Chromosoma 81, 65–84 (1980). https://doi.org/10.1007/BF00292423

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00292423

Keywords

Search

Navigation