Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1985 Dec 30;4(13B):3757–3764. doi: 10.1002/j.1460-2075.1985.tb04145.x

Cloning of the homeotic Sex combs reduced gene in Drosophila and in situ localization of its transcripts

Atsushi Kuroiwa 1,1, Urs Kloter 1, Paul Baumgartner 1, Walter J Gehring 1
PMCID: PMC554728  PMID: 16453653

Abstract

We have extended our `chromosomal walk' in the Antennapedia-complex (ANT-C) by isolating overlapping DNA sequences spanning the chromosomal segment between Antennapedia (Antp) and Deformed (Dfd). The transcription units, homeoboxes and M-repeats were mapped within this region. Four transcription units Antp, fushi tarazu (ftz), Sex combs reduced (Scr) and Dfd contain both a homeobox and an M repeat, whereas at least two additional transcription units, x and z, were found to lack these elements. The Scr locus was identified by deletion mapping. It consists of at least two exonic regions separated by a large intron. The homeobox is located in the 3' exon and is 82% homologous to the one in Antp. Scr encodes a major 3.9-kb RNA. A corresponding cDNA clone was used as a probe for in situ hybridization to sections of various embryonic stages. At gastrula stages Scr+ transcripts accumulate in the posterior head and the anterior thoracic region of the germ band. At later stages a strong accumulation of transcripts is observed in the suboesophageal and the prothoracic ganglion of the ventral nervous system.

Keywords: Drosophila, homeotic genes, Sex combs reduced, cloning, homeobox

Full text

PDF
3762

Images in this article

3758Fig. 2.
on p.3758
3759Fig. 3.
on p.3759
3760Fig. 4.
on p.3760
3760Fig. 6.
on p.3760
3762Fig. 8.
on p.3762
3763Fig. 9.
on p.3763

Selected References

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

  1. Anderson K. V., Nüsslein-Volhard C. Information for the dorsal--ventral pattern of the Drosophila embryo is stored as maternal mRNA. Nature. 1984 Sep 20;311(5983):223–227. doi: 10.1038/311223a0. [DOI] [PubMed] [Google Scholar]
  2. Beachy P. A., Helfand S. L., Hogness D. S. Segmental distribution of bithorax complex proteins during Drosophila development. Nature. 1985 Feb 14;313(6003):545–551. doi: 10.1038/313545a0. [DOI] [PubMed] [Google Scholar]
  3. Garber R. L., Kuroiwa A., Gehring W. J. Genomic and cDNA clones of the homeotic locus Antennapedia in Drosophila. EMBO J. 1983;2(11):2027–2036. doi: 10.1002/j.1460-2075.1983.tb01696.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gehring W. J. The homeo box: a key to the understanding of development? Cell. 1985 Jan;40(1):3–5. doi: 10.1016/0092-8674(85)90300-9. [DOI] [PubMed] [Google Scholar]
  5. Hafen E., Levine M., Garber R. L., Gehring W. J. An improved in situ hybridization method for the detection of cellular RNAs in Drosophila tissue sections and its application for localizing transcripts of the homeotic Antennapedia gene complex. EMBO J. 1983;2(4):617–623. doi: 10.1002/j.1460-2075.1983.tb01472.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hazelrigg T., Kaufman T. C. Revertants of Dominant Mutations Associated with the Antennapedia Gene Complex of DROSOPHILA MELANOGASTER: Cytology and Genetics. Genetics. 1983 Nov;105(3):581–600. doi: 10.1093/genetics/105.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kaufman T. C., Lewis R., Wakimoto B. Cytogenetic Analysis of Chromosome 3 in DROSOPHILA MELANOGASTER: The Homoeotic Gene Complex in Polytene Chromosome Interval 84a-B. Genetics. 1980 Jan;94(1):115–133. doi: 10.1093/genetics/94.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kuroiwa A., Hafen E., Gehring W. J. Cloning and transcriptional analysis of the segmentation gene fushi tarazu of Drosophila. Cell. 1984 Jul;37(3):825–831. doi: 10.1016/0092-8674(84)90417-3. [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. Lepesant J. A., Kejzlarova-Lepesant J., Garen A. Ecdysone-inducible functions of larval fat bodies in Drosophila. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5570–5574. doi: 10.1073/pnas.75.11.5570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lewis E. B. A gene complex controlling segmentation in Drosophila. Nature. 1978 Dec 7;276(5688):565–570. doi: 10.1038/276565a0. [DOI] [PubMed] [Google Scholar]
  12. Lewis R. A., Kaufman T. C., Denell R. E., Tallerico P. Genetic Analysis of the Antennapedia Gene Complex (Ant-C) and Adjacent Chromosomal Regions of DROSOPHILA MELANOGASTER. I. Polytene Chromosome Segments 84b-D. Genetics. 1980 Jun;95(2):367–381. doi: 10.1093/genetics/95.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lewis R. A., Wakimoto B. T., Denell R. E., Kaufman T. C. Genetic Analysis of the Antennapedia Gene Complex (Ant-C) and Adjacent Chromosomal Regions of DROSOPHILA MELANOGASTER. II. Polytene Chromosome Segments 84A-84B1,2. Genetics. 1980 Jun;95(2):383–397. doi: 10.1093/genetics/95.2.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  15. McGinnis W., Garber R. L., Wirz J., Kuroiwa A., Gehring W. J. A homologous protein-coding sequence in Drosophila homeotic genes and its conservation in other metazoans. Cell. 1984 Jun;37(2):403–408. doi: 10.1016/0092-8674(84)90370-2. [DOI] [PubMed] [Google Scholar]
  16. McGinnis W., Levine M. S., Hafen E., Kuroiwa A., Gehring W. J. A conserved DNA sequence in homoeotic genes of the Drosophila Antennapedia and bithorax complexes. 1984 Mar 29-Apr 4Nature. 308(5958):428–433. doi: 10.1038/308428a0. [DOI] [PubMed] [Google Scholar]
  17. Medford R. M., Nguyen H. T., Destree A. T., Summers E., Nadal-Ginard B. A novel mechanism of alternative RNA splicing for the developmentally regulated generation of troponin T isoforms from a single gene. Cell. 1984 Sep;38(2):409–421. doi: 10.1016/0092-8674(84)90496-3. [DOI] [PubMed] [Google Scholar]
  18. Morrison D. A. Transformation and preservation of competent bacterial cells by freezing. Methods Enzymol. 1979;68:326–331. doi: 10.1016/0076-6879(79)68023-0. [DOI] [PubMed] [Google Scholar]
  19. Nüsslein-Volhard C., Wieschaus E. Mutations affecting segment number and polarity in Drosophila. Nature. 1980 Oct 30;287(5785):795–801. doi: 10.1038/287795a0. [DOI] [PubMed] [Google Scholar]
  20. Ouweneel W. J. Developmental genetics of homoeosis. Adv Genet. 1976;18:179–248. doi: 10.1016/s0065-2660(08)60439-3. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Scott M. P., Weiner A. J., Hazelrigg T. I., Polisky B. A., Pirrotta V., Scalenghe F., Kaufman T. C. The molecular organization of the Antennapedia locus of Drosophila. Cell. 1983 Dec;35(3 Pt 2):763–776. doi: 10.1016/0092-8674(83)90109-5. [DOI] [PubMed] [Google Scholar]
  23. Scott M. P., Weiner A. J. Structural relationships among genes that control development: sequence homology between the Antennapedia, Ultrabithorax, and fushi tarazu loci of Drosophila. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4115–4119. doi: 10.1073/pnas.81.13.4115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Struhl G. A homoeotic mutation transforming leg to antenna in Drosophila. Nature. 1981 Aug 13;292(5824):635–638. doi: 10.1038/292635a0. [DOI] [PubMed] [Google Scholar]
  26. Struhl G. Role of the esc+ gene product in ensuring the selective expression of segment-specific homeotic genes in Drosophila. J Embryol Exp Morphol. 1983 Aug;76:297–331. [PubMed] [Google Scholar]
  27. Sutcliffe J. G., Milner R. J., Gottesfeld J. M., Reynolds W. Control of neuronal gene expression. Science. 1984 Sep 21;225(4668):1308–1315. doi: 10.1126/science.6474179. [DOI] [PubMed] [Google Scholar]
  28. Sánchez-Herrero E., Vernós I., Marco R., Morata G. Genetic organization of Drosophila bithorax complex. Nature. 1985 Jan 10;313(5998):108–113. doi: 10.1038/313108a0. [DOI] [PubMed] [Google Scholar]
  29. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  30. Wakimoto B. T., Kaufman T. C. Analysis of larval segmentation in lethal genotypes associated with the antennapedia gene complex in Drosophila melanogaster. Dev Biol. 1981 Jan 15;81(1):51–64. doi: 10.1016/0012-1606(81)90347-x. [DOI] [PubMed] [Google Scholar]
  31. Wakimoto B. T., Turner F. R., Kaufman T. C. Defects in embryogenesis in mutants associated with the antennapedia gene complex of Drosophila melanogaster. Dev Biol. 1984 Mar;102(1):147–172. doi: 10.1016/0012-1606(84)90182-9. [DOI] [PubMed] [Google Scholar]
  32. Weiner A. J., Scott M. P., Kaufman T. C. A molecular analysis of fushi tarazu, a gene in Drosophila melanogaster that encodes a product affecting embryonic segment number and cell fate. Cell. 1984 Jul;37(3):843–851. doi: 10.1016/0092-8674(84)90419-7. [DOI] [PubMed] [Google Scholar]
  33. Wharton K. A., Yedvobnick B., Finnerty V. G., Artavanis-Tsakonas S. opa: a novel family of transcribed repeats shared by the Notch locus and other developmentally regulated loci in D. melanogaster. Cell. 1985 Jan;40(1):55–62. doi: 10.1016/0092-8674(85)90308-3. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES