Chromosoma

, Volume 31, Issue 3, pp 331–345 | Cite as

Localisation of reiterated nucleotide sequences in Drosophila and mouse by in situ hybridisation of complementary RNA

  • Kenneth W. Jones
  • Forbes W. Robertson
Article

Abstract

The location of highly reiterated nucleotide sequences on the chromosomes has been studied by the technique of in situ hybridisation between the DNA of either Drosophila melanogaster salivary gland chromosomes or mouse chromosomes and tritium labelled complementary RNA (c-RNA) transcribed in vitro from appropriate templates with the aid of DNA dependent RNA polymerase extracted from Micrococcus lysodeikticus. The location of the hybrid material was identified by autoradiography after RNase treatment. — When Drosophila c-RNA, transcribed from whole DNA, was annealed with homologous salivary chromosomes in the presence of formamide the well defined labelling was confined to the chromocentre. With heat instead of formamide denaturation there was evidence of discontinuous labelling in various chromosome regions as well, apparently associated with banding. Xenopus ribosomal RNA showed no evidence of annealing to Drosophila chromosomes with the comparatively short exposure times used here. — When mouse satellite DNA was used as template the resulting c-RNA showed no hybridisation to Drosophila chromosomes but, when annealed with mouse chromosomes, the centromeric regions were intensely labelled. The interphase nuclei showed several distinct regions of high activity which suggested aggregation of centromeric regions of both homologous and non-homologous chromosomes. The results of annealing either c-RNA or labelled satellite DNA to homologous chromosomes were virtually indistinguishable. Incubation of Drosophila c-RNA with mouse chromosomes provided no evidence of localisation of grains. — It is inferred that both in mouse and Drosophila the centromeric regions of all chromosomes are enriched in highly reiterated sequences. This may be a general phenomenon and it might be tentatively suggested that the highly reiterated sequences play some role in promoting the close physical approximation of homologous and non-homologous chromosomes or chromosome regions to facilitate regulation of function.

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References

  1. Bishop, J. C., Robertson, F. W.: Transcription of bacteriophage T4 deoxyribo-nucleic acid in vitro. Biochem. J. 115, 353–370 (1969).Google Scholar
  2. Brown, D. D., Weber, C. S., Sinclair, J. H.: Ribosomal RNA and its genes during oogenesis and development. Carnegie Inst. Yearbook 66, 580–589 (1967).Google Scholar
  3. Flamm, W. G., Bond, H. E., Burr, H. B.: Density gradient centrifugation in a fixed angle rotor; a higher order of resolution. Biochem. biophys. Acta (Amst.) 129, 310–317 (1968).Google Scholar
  4. Gall, J. G., Pardue, M. L.: Formation and detection of RNA-DNA hybrid molecules in cytological preparation. Proc. nat. Acad. Sci. (Wash.) 63, 378–383 (1969).Google Scholar
  5. Harel, J., Hanania, N., Tapiero, H., Harel, L.: RNA replication by nuclear satellite DNA in different mouse cells. Biochem. biophys. Res. Commun. 33, 696–701 (1968).Google Scholar
  6. John, H. A., Birnstiel, M., Jones, K. W.: RNA/DNA hybrids at the cytological level. Nature (Lond.) 223, 582–587 (1969).Google Scholar
  7. Jones, K. W.: The chromosomal and nuclear location of mouse satellite DNA in individual cells. Nature (Lond.) 225, 912–915 (1970).Google Scholar
  8. Macgregor, H. C.: Nucleolar DNA in oooytes of Xenopus laevis. J. Cell Sci. 3, 437–444 (1968).Google Scholar
  9. McConaughy, B. L., Laird, C. D., McCarthy, B. J.: Nucleic acid reassociation in formamide. Biochemistry 8, 3289–3295 (1969).Google Scholar
  10. Nakamoto, T., Fox, C. F., Weiss, S. B.: Enzymatic synthesis of ribonucleic acid 1. Preparation of ribonucleic acid polymerase from extracts of Micrococcus lysodeikticus. J. biol. Chem. 239, 167–174 (1964).Google Scholar
  11. Pardue, M. L., Gall, J. G.: Molecular hybridisation of radioactive DNA to the DNA of cytological preparations. Proc. nat. Acad. Sci. (Wash.) 64, 600–604 (1969).Google Scholar
  12. —, Gerbi, S. A., Eckhardt, R. A., Gall, J. C.: Cytological localisation of DNA complementary to ribosomal RNA in polytene chromosomes of Diptera. Chromosoma (Berl.) 29, 268–290 (1970).Google Scholar
  13. Robertson, F. W., Chipchase, M., Nguyen, T. M.: The comparison of differences in reiterated sequences by RNA-DNA hybridisation. Genetics 63, 369–385 (1969).Google Scholar
  14. Schildkraut, K. L., Maio, J. J.: Studies on the intranucleic distribution and properties of mouse satellite DNA. Biochim. biophys. Acta (Amst.) 161, 76–93 (1968).Google Scholar
  15. Southern, E.: In preparation (1970).Google Scholar
  16. Walker, P. M. B., Flamm, W. G., McLaren, A.: Highly repetitive DNA in nuclei. Handbook of molecular cytology (Lima-de-Faria, ed.), p. 52–66. Amsterdam: North Holland Publ. Co. 1969.Google Scholar

Copyright information

© Springer-Verlag 1970

Authors and Affiliations

  • Kenneth W. Jones
    • 1
  • Forbes W. Robertson
    • 1
    • 2
  1. 1.Department of GeneticsUniversity of EdinburghScotland
  2. 2.Department of GeneticsUniversity of AberdeenAberdeenScotland

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