Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Jul 1;99(1 Pt 1):233–238. doi: 10.1083/jcb.99.1.233

A cytological approach to the ordering of events in gene activation using the Sgs-4 locus of Drosophila melanogaster

PMCID: PMC2275650  PMID: 6330126

Abstract

The polytene chromosomes of Drosophila strains that differ in the synthesis of the major salivary gland glue protein sgs-4 were examined by indirect immunofluorescence using antisera to several nonhistone chromosomal proteins. The Oregon-R X chromosome, which produces sgs-4 messenger RNA, showed a strong fluorescent band at locus 3C11-12 when stained with anti-RNA polymerase II, whereas the null mutant Berkeley 1 failed to exhibit fluorescence at that locus. The presence of another antigen (Band 2), normally associated with developmentally active loci, was clearly evident at locus 3C11-12 of both transcriptionally competent and null strains, indicating that the association of Band 2 antigen with the chromatin is an event independent of RNA polymerase II binding. Antibodies directed against Drosophila topoisomerase I stained 3C11-12 in the Sgs-4+ (wild-type) strain brightly, but gave significantly less staining in the null strain. This indicates that the high concentrations of topoisomerase I seen at active loci are closely associated with the transcriptional event. In some of these analyses, we have made use of flies heterozygous for the wild-type and null alleles in order to make internally controlled comparisons. The results suggest that this type of analysis will enable conclusions to be drawn concerning the interdependence and order of action of chromosomal proteins involved in developmental gene activation.

Full Text

The Full Text of this article is available as a PDF (738.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alfageme C. R., Rudkin G. T., Cohen L. H. Locations of chromosomal proteins in polytene chromosomes. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2038–2042. doi: 10.1073/pnas.73.6.2038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beckendorf S. K., Kafatos F. C. Differentiation in the salivary glands of Drosophila melanogaster: characterization of the glue proteins and their developmental appearance. Cell. 1976 Nov;9(3):365–373. doi: 10.1016/0092-8674(76)90081-7. [DOI] [PubMed] [Google Scholar]
  3. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  4. Elgin S. C. DNAase I-hypersensitive sites of chromatin. Cell. 1981 Dec;27(3 Pt 2):413–415. doi: 10.1016/0092-8674(81)90381-0. [DOI] [PubMed] [Google Scholar]
  5. Jamrich M., Greenleaf A. L., Bautz E. K. Localization of RNA polymerase in polytene chromosomes of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1977 May;74(5):2079–2083. doi: 10.1073/pnas.74.5.2079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kabisch R., Bautz E. K. Differential distribution of RNA polymerase B and nonhistone chromosomal proteins in polytene chromosomes of Drosophila melanogaster. EMBO J. 1983;2(3):395–402. doi: 10.1002/j.1460-2075.1983.tb01436.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Korge G. Chromosome puff activity and protein synthesis in larval salivary glands of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4550–4554. doi: 10.1073/pnas.72.11.4550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Korge G. Genetic analysis of the larval secretion gene Sgs-4 and its regulatory chromosome sites in Drosophila melanogaster. Chromosoma. 1981;84(3):373–390. doi: 10.1007/BF00286027. [DOI] [PubMed] [Google Scholar]
  9. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  10. Lowenhaupt K., Cartwright I. L., Keene M. A., Zimmerman J. L., Elgin S. C. Chromatin structure in pre- and postblastula embryos of Drosophila. Dev Biol. 1983 Sep;99(1):194–201. doi: 10.1016/0012-1606(83)90267-1. [DOI] [PubMed] [Google Scholar]
  11. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mayfield J. E., Serunian L. A., Silver L. M., Elgin S. C. A protein released by DNAase I digestion of drosophila nuclei is preferentially associated with puffs. Cell. 1978 Jul;14(3):539–544. doi: 10.1016/0092-8674(78)90240-4. [DOI] [PubMed] [Google Scholar]
  13. McGinnis W., Farrell J., Jr, Beckendorf S. K. Molecular limits on the size of a genetic locus in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7367–7371. doi: 10.1073/pnas.77.12.7367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McGinnis W., Shermoen A. W., Beckendorf S. K. A transposable element inserted just 5' to a Drosophila glue protein gene alters gene expression and chromatin structure. Cell. 1983 Aug;34(1):75–84. doi: 10.1016/0092-8674(83)90137-x. [DOI] [PubMed] [Google Scholar]
  15. Meyerowitz E. M., Hogness D. S. Molecular organization of a Drosophila puff site that responds to ecdysone. Cell. 1982 Jan;28(1):165–176. doi: 10.1016/0092-8674(82)90386-5. [DOI] [PubMed] [Google Scholar]
  16. Muskavitch M. A., Hogness D. S. An expandable gene that encodes a Drosophila glue protein is not expressed in variants lacking remote upstream sequences. Cell. 1982 Jul;29(3):1041–1051. doi: 10.1016/0092-8674(82)90467-6. [DOI] [PubMed] [Google Scholar]
  17. Muskavitch M. A., Hogness D. S. Molecular analysis of a gene in a developmentally regulated puff of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7362–7366. doi: 10.1073/pnas.77.12.7362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Plagens U., Greenleaf A. L., Bautz E. K. Distribution of RNA polymerase on Drosophila polytene chromosomes as studied by indirect immunofluorescence. Chromosoma. 1976 Dec 16;59(2):157–165. doi: 10.1007/BF00328484. [DOI] [PubMed] [Google Scholar]
  19. Rubin G. M., Spradling A. C. Genetic transformation of Drosophila with transposable element vectors. Science. 1982 Oct 22;218(4570):348–353. doi: 10.1126/science.6289436. [DOI] [PubMed] [Google Scholar]
  20. Shermoen A. W., Beckendorf S. K. A complex of interacting DNAase I-hypersensitive sites near the Drosophila glue protein gene, Sgs4. Cell. 1982 Jun;29(2):601–607. doi: 10.1016/0092-8674(82)90176-3. [DOI] [PubMed] [Google Scholar]
  21. Silver L. M., Elgin S. C. A method for determination of the in situ distribution of chromosomal proteins. Proc Natl Acad Sci U S A. 1976 Feb;73(2):423–427. doi: 10.1073/pnas.73.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Weeks J. R., Coulter D. E., Greenleaf A. L. Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme. J Biol Chem. 1982 May 25;257(10):5884–5892. [PubMed] [Google Scholar]
  24. Weintraub H., Beug H., Groudine M., Graf T. Temperature-sensitive changes in the structure of globin chromatin in lines of red cell precursors transformed by ts-AEV. Cell. 1982 Apr;28(4):931–940. doi: 10.1016/0092-8674(82)90072-1. [DOI] [PubMed] [Google Scholar]
  25. Wu C. The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature. 1980 Aug 28;286(5776):854–860. doi: 10.1038/286854a0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES