Chromosoma

, Volume 86, Issue 1, pp 77–93 | Cite as

Interbands of polytene chromosomes: Binding sites and start points for RNA polymerase B (II)

  • Heinz Sass
  • Ekkehard K. F. Bautz
Article

Abstract

Polytene chromosomes of different chironomids, i.e., Chironomus tentans, C. melanotus and Glyptotendipes barbipes were isolated from salivary glands in a native state. These chromosomes were treated in vitro either mechanically or with different ionic strengths to modify them structurally as to yield different degrees of decondensation of the compact bands. Treated and untreated polytene chromosomes were lightly fixed with formaldehyde and stained by indirect immunofluorescence for RNA polymerase B. The distribution of this enzyme in bands, interbands, puffs and centromeric heterochromatin was scored and compared with that of histone H2B. The results indicate that failure to observe an antigen in condensed regions of chromatin does not necessarily imply its absence. Decondensation of bands, for example, leads to massive uncovering of histone H2B antigen, which appears to be masked in the bands of untreated polytene chromosomes. No evidence, however, of a corresponding unmasking of RNA polymerase B molecules was observed, indicating that few if any enzyme molecules are trapped in bands. Thus binding sites for RNA polymerase B and start points for transcriptional activity of the enzyme appear always to be the interband regions.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Alfageme, C.R., Rudkin, G.T., Cohen, L.H.: Locations of chromosomal proteins in polytene chromosomes. Proc. Natl. Acad. Sci. (Wash.) 73, 2038–2042 (1976)Google Scholar
  2. Ananiev, E.B., 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
  3. Beermann, W.: Chromomerenkonstanz und spezifische Modifikation der Chromosomenstruktur in der Entwicklung und Organdifferenzierung von Chironomus tentans. Chromosoma (Berl.) 5, 139–198 (1952)Google Scholar
  4. Beermann, W.: Cytologische Analyse eines Camptochironomus-Artbastards. I. Kreuzungsergebnisse und die Evolution des Karyotypus. Chromosoma (Berl.) 7, 198–259 (1955)Google Scholar
  5. Berendes, H.D.: Factors involved in the expression of gene activity in polytene chromosomes. Chromosoma (Berl.) 24, 418–437 (1968)Google Scholar
  6. Bustin, M., Kurth, P.D., Moudrianakis, E.N., Goldblatt, D., Sperling, R., Rizzo, W.B.: Immunological probes for chromatin structure. Cold Spr. Harb. Sym. quant. Biol. 42, 379–388 (1978)Google Scholar
  7. Greenleaf, A.L., Bautz, E.K.F.: RNA polymerase B from Drosophila melanogaster larvae. Purification and partial characterization. Europ. J. Biochem. 60, 169–179 (1975)Google Scholar
  8. Hameister, H.: RNA synthesis in isolated polytene nuclei from Chironomus tentans. Chromosoma (Berl.) 62, 217–242 (1977)Google Scholar
  9. Holt, T.K.H.: Local protein accumulation during gene activation. I. Quantitative measurements on dye binding capacity at subsequent stages of puff formation in D. hydei. Chromosoma (Berl.) 32, 64–78 (1970)Google Scholar
  10. Hsieh, T., Brutlag, D.L.: A protein that preferentially binds Drosophila satellite DNA. Proc. Natl. Acad. Sci. (Wash.) 76, 726–730 (1979)Google Scholar
  11. Jamrich, M., Greenleaf, A.L., Bautz, E.K.F.: Localization of RNA polymerase in polytene chromosomes of Drosophila melanogaster. Proc. Natl. Acad. Sci. (Wash.) 74, 2079–2083 (1977)Google Scholar
  12. Krämer, A., Haars, R., Kabisch, R., Will, H., Bautz, F.A., Bautz, E.K.F.: Monoclonal antibody directed against RNA polymerase II of Drosophila melanogaster. Molec. Gen. Genet. 180, 193–199 (1980)Google Scholar
  13. Lakhotia, S.C., Jacob, J.: EM autoradiographic studies on polytene nuclei of Drosophila melanogaster. II. Organization and transcriptive activity of the chromocentre. Exp. Cell Res. 86, 253–263 (1974)Google Scholar
  14. Lezzi, M., Robert, M.: Chromosomes isolated from unfixed salivary glands of Chironomus. In: Results and problems in cell differentiation (W. Beermann, ed.), vol. 4, pp. 35–47. Berlin, Heidelberg, New York: Springer-Verlag 1972Google Scholar
  15. Lezzi, M., Meyer, B., Mähr, R.: Heat shock phenomena in Chironomus tentans. I. In vivo effects of heat, overheat, and quenching on salivary chromosome puffing. Chromosoma (Berl.) 83, 327–339 (1981)Google Scholar
  16. Pelling, C.: Ribonukleinsäure-Synthese der Riesenchromosomen. Autoradiographische Untersuchungen an Chironomus tentans. Chromosoma (Berl.) 15, 71–122 (1964)Google Scholar
  17. Sass, H.: Features of in vitro puffing and RNA synthesis in polytene chromosomes of Chironomus. Chromosoma (Berl.) 78, 33–78 (1980a)Google Scholar
  18. Sass, H.: Hierarchy of fibrillar organization levels in the polytene interphase chromosomes of Chironomus. J. Cell Sci. 45, 269–293 (1980b)Google Scholar
  19. Sass, H.: Effects of DMSO on the structure and function of polytene chromosomes of Chironomus. Chromosoma (Berl.) 83, 619–643 (1981)Google Scholar
  20. Sass, H.: RNA polymerase B in polytene chromosomes: Immunofluorescent and autoradiographic analysis during stimulated and repressed RNA synthesis. Cell 28, 269–278 (1982)Google Scholar
  21. Sass, H., Bautz, E.K.F.: Immunoelectron microscopic localization of RNA polymerase B on isolated polytene chromosomes of Chironomus tentans. Chromosoma (Berl.) 85, 633–642 (1982)Google Scholar
  22. Schmidt, E.R.: Two AT-rich satellite DNAs in the chironomid Glyptotendipes barbipes (Staeger). Isolation and localization in polytene chromosomes of G. barbipes and Chironomus thummi. Chromosoma (Berl.) 79, 315–328 (1980)Google Scholar
  23. Steinemann, M.: Co-replication of satellite DNA of Chironomus melanotus with mainband DNA during polytenization. Chromosoma (Berl.) 66, 127–139 (1978)Google Scholar
  24. Walter, L.: Syntheseprozesse an den Riesenchromosomen von Glyptotendipes. Chromosoma (Berl.) 41, 327–360 (1973)Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Heinz Sass
    • 1
  • Ekkehard K. F. Bautz
    • 1
  1. 1.Molekulare Genetik der Universität HeidelbergHeidelbergGermany

Personalised recommendations