Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 2001 Feb;157(2):679–688. doi: 10.1093/genetics/157.2.679

Molecular identification of virilizer, a gene required for the expression of the sex-determining gene Sex-lethal in Drosophila melanogaster.

M Niessen 1, R Schneiter 1, R Nothiger 1
PMCID: PMC1461513  PMID: 11156988

Abstract

Sex-lethal (Sxl) is a central switch gene in somatic sexual development of Drosophila melanogaster. Female-specific expression of Sxl relies on autoregulatory splicing of Sxl pre-mRNA by SXL protein. This process requires the function of virilizer (vir). Besides its role in Sxl splicing, vir is essential for male and female viability and is also required for the production of eggs capable of embryonic development. We have identified vir molecularly and found that it produces a single transcript of 6 kb that is ubiquitously expressed in male and female embryos throughout development. This transcript encodes a nuclear protein of 210 kD that cannot be assigned to a known protein family. VIR contains a putative transmembrane domain, a coiled-coil region and PEST sequences. We have characterized five different alleles of vir. Those alleles that affect both sexes are associated with large truncations of the protein, while alleles that affect only the female-specific functions are missense mutations that lie relatively close to each other, possibly defining a region important for the regulation of Sxl.

Full Text

The Full Text of this article is available as a PDF (379.1 KB).

Selected References

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

  1. Albrecht E. B., Salz H. K. The Drosophila sex determination gene snf is utilized for the establishment of the female-specific splicing pattern of Sex-lethal. Genetics. 1993 Jul;134(3):801–807. doi: 10.1093/genetics/134.3.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bashaw G. J., Baker B. S. The regulation of the Drosophila msl-2 gene reveals a function for Sex-lethal in translational control. Cell. 1997 May 30;89(5):789–798. doi: 10.1016/s0092-8674(00)80262-7. [DOI] [PubMed] [Google Scholar]
  3. Bopp D., Bell L. R., Cline T. W., Schedl P. Developmental distribution of female-specific Sex-lethal proteins in Drosophila melanogaster. Genes Dev. 1991 Mar;5(3):403–415. doi: 10.1101/gad.5.3.403. [DOI] [PubMed] [Google Scholar]
  4. Burnette J. M., Hatton A. R., Lopez A. J. Trans-acting factors required for inclusion of regulated exons in the Ultrabithorax mRNAs of Drosophila melanogaster. Genetics. 1999 Apr;151(4):1517–1529. doi: 10.1093/genetics/151.4.1517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Cline T. W., Meyer B. J. Vive la différence: males vs females in flies vs worms. Annu Rev Genet. 1996;30:637–702. doi: 10.1146/annurev.genet.30.1.637. [DOI] [PubMed] [Google Scholar]
  7. Corpet F., Gouzy J., Kahn D. Recent improvements of the ProDom database of protein domain families. Nucleic Acids Res. 1999 Jan 1;27(1):263–267. doi: 10.1093/nar/27.1.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Flickinger T. W., Salz H. K. The Drosophila sex determination gene snf encodes a nuclear protein with sequence and functional similarity to the mammalian U1A snRNP protein. Genes Dev. 1994 Apr 15;8(8):914–925. doi: 10.1101/gad.8.8.914. [DOI] [PubMed] [Google Scholar]
  10. Frischauf A. M., Lehrach H., Poustka A., Murray N. Lambda replacement vectors carrying polylinker sequences. J Mol Biol. 1983 Nov 15;170(4):827–842. doi: 10.1016/s0022-2836(83)80190-9. [DOI] [PubMed] [Google Scholar]
  11. Gebauer F., Merendino L., Hentze M. W., Valcárcel J. The Drosophila splicing regulator sex-lethal directly inhibits translation of male-specific-lethal 2 mRNA. RNA. 1998 Feb;4(2):142–150. [PMC free article] [PubMed] [Google Scholar]
  12. Granadino B., Campuzano S., Sánchez L. The Drosophila melanogaster fl(2)d gene is needed for the female-specific splicing of Sex-lethal RNA. EMBO J. 1990 Aug;9(8):2597–2602. doi: 10.1002/j.1460-2075.1990.tb07441.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Granadino B., San Juán A., Santamaria P., Sánchez L. Evidence of a dual function in fl(2)d, a gene needed for Sex-lethal expression in Drosophila melanogaster. Genetics. 1992 Mar;130(3):597–612. doi: 10.1093/genetics/130.3.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horabin J. I., Schedl P. Regulated splicing of the Drosophila sex-lethal male exon involves a blockage mechanism. Mol Cell Biol. 1993 Mar;13(3):1408–1414. doi: 10.1128/mcb.13.3.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Horabin J. I., Schedl P. Sex-lethal autoregulation requires multiple cis-acting elements upstream and downstream of the male exon and appears to depend largely on controlling the use of the male exon 5' splice site. Mol Cell Biol. 1993 Dec;13(12):7734–7746. doi: 10.1128/mcb.13.12.7734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Inoue K., Hoshijima K., Sakamoto H., Shimura Y. Binding of the Drosophila sex-lethal gene product to the alternative splice site of transformer primary transcript. Nature. 1990 Mar 29;344(6265):461–463. doi: 10.1038/344461a0. [DOI] [PubMed] [Google Scholar]
  17. Kelley R. L., Wang J., Bell L., Kuroda M. I. Sex lethal controls dosage compensation in Drosophila by a non-splicing mechanism. Nature. 1997 May 8;387(6629):195–199. doi: 10.1038/387195a0. [DOI] [PubMed] [Google Scholar]
  18. Keyes L. N., Cline T. W., Schedl P. The primary sex determination signal of Drosophila acts at the level of transcription. Cell. 1992 Mar 6;68(5):933–943. doi: 10.1016/0092-8674(92)90036-c. [DOI] [PubMed] [Google Scholar]
  19. Lupas A. Coiled coils: new structures and new functions. Trends Biochem Sci. 1996 Oct;21(10):375–382. [PubMed] [Google Scholar]
  20. Lupas A., Van Dyke M., Stock J. Predicting coiled coils from protein sequences. Science. 1991 May 24;252(5009):1162–1164. doi: 10.1126/science.252.5009.1162. [DOI] [PubMed] [Google Scholar]
  21. Ma K., Inglis J. D., Sharkey A., Bickmore W. A., Hill R. E., Prosser E. J., Speed R. M., Thomson E. J., Jobling M., Taylor K. A Y chromosome gene family with RNA-binding protein homology: candidates for the azoospermia factor AZF controlling human spermatogenesis. Cell. 1993 Dec 31;75(7):1287–1295. doi: 10.1016/0092-8674(93)90616-x. [DOI] [PubMed] [Google Scholar]
  22. O'Connell P. O., Rosbash M. Sequence, structure, and codon preference of the Drosophila ribosomal protein 49 gene. Nucleic Acids Res. 1984 Jul 11;12(13):5495–5513. doi: 10.1093/nar/12.13.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Penalva L. O., Ruiz M. F., Ortega A., Granadino B., Vicente L., Segarra C., Valcárcel J., Sánchez L. The Drosophila fl(2)d gene, required for female-specific splicing of Sxl and tra pre-mRNAs, encodes a novel nuclear protein with a HQ-rich domain. Genetics. 2000 May;155(1):129–139. doi: 10.1093/genetics/155.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Reardon J. T., Liljestrand-Golden C. A., Dusenbery R. L., Smith P. D. Molecular Analysis of Diepoxybutane-Induced Mutations at the rosy Locus of Drosophila melanogaster. Genetics. 1987 Feb;115(2):323–331. doi: 10.1093/genetics/115.2.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sakamoto H., Inoue K., Higuchi I., Ono Y., Shimura Y. Control of Drosophila Sex-lethal pre-mRNA splicing by its own female-specific product. Nucleic Acids Res. 1992 Nov 11;20(21):5533–5540. doi: 10.1093/nar/20.21.5533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Samuels M., Deshpande G., Schedl P. Activities of the Sex-lethal protein in RNA binding and protein:protein interactions. Nucleic Acids Res. 1998 Jun 1;26(11):2625–2637. doi: 10.1093/nar/26.11.2625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schütt C., Hilfiker A., Nöthiger R. virilizer regulates Sex-lethal in the germline of Drosophila melanogaster. Development. 1998 Apr;125(8):1501–1507. doi: 10.1242/dev.125.8.1501. [DOI] [PubMed] [Google Scholar]
  28. Sosnowski B. A., Belote J. M., McKeown M. Sex-specific alternative splicing of RNA from the transformer gene results from sequence-dependent splice site blockage. Cell. 1989 Aug 11;58(3):449–459. doi: 10.1016/0092-8674(89)90426-1. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Stitzinger S. M., Pellicena-Palle A., Albrecht E. B., Gajewski K. M., Beckingham K. M., Salz H. K. Mutations in the predicted aspartyl tRNA synthetase of Drosophila are lethal and function as dosage-sensitive maternal modifiers of the sex determination gene Sex-lethal. Mol Gen Genet. 1999 Feb;261(1):142–151. doi: 10.1007/s004380050951. [DOI] [PubMed] [Google Scholar]
  31. Thisse B., el Messal M., Perrin-Schmitt F. The twist gene: isolation of a Drosophila zygotic gene necessary for the establishment of dorsoventral pattern. Nucleic Acids Res. 1987 Apr 24;15(8):3439–3453. doi: 10.1093/nar/15.8.3439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wahl G. M., Lewis K. A., Ruiz J. C., Rothenberg B., Zhao J., Evans G. A. Cosmid vectors for rapid genomic walking, restriction mapping, and gene transfer. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2160–2164. doi: 10.1073/pnas.84.8.2160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Welihinda A. A., Tirasophon W., Kaufman R. J. The cellular response to protein misfolding in the endoplasmic reticulum. Gene Expr. 1999;7(4-6):293–300. [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES