Skip to main content
Genetics logoLink to Genetics
. 2000 May;155(1):129–139. doi: 10.1093/genetics/155.1.129

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.

L O Penalva 1, M F Ruiz 1, A Ortega 1, B Granadino 1, L Vicente 1, C Segarra 1, J Valcárcel 1, L Sánchez 1
PMCID: PMC1461084  PMID: 10790389

Abstract

The Drosophila gene female-lethal(2)d [fl(2)d] interacts genetically with the master regulatory gene for sex determination, Sex-lethal. Both genes are required for the activation of female-specific patterns of alternative splicing on transformer and Sex-lethal pre-mRNAs. We have used P-element-mediated mutagenesis to identify the fl(2)d gene. The fl(2)d transcription unit generates two alternatively spliced mRNAs that can encode two protein isoforms differing at their amino terminus. The larger isoform contains a domain rich in histidine and glutamine but has no significant homology to proteins in databases. Several lines of evidence indicate that this protein is responsible for fl(2)d function. First, the P-element insertion that inactivates fl(2)d interrupts this ORF. Second, amino acid changes within this ORF have been identified in fl(2)d mutants, and the nature of the changes correlates with the severity of the mutations. Third, all of the phenotypes associated with fl(2)d mutations can be rescued by expression of this cDNA in transgenic flies. Fl(2)d protein can be detected in extracts from Drosophila cell lines, embryos, larvae, and adult animals, without apparent differences between sexes, as well as in adult ovaries. Consistent with a possible function in posttranscriptional regulation, Fl(2)d protein has nuclear localization and is enriched in nuclear extracts.

Full Text

The Full Text of this article is available as a PDF (322.7 KB).

Selected References

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

  1. Alkhatib G., Briedis D. J. The predicted primary structure of the measles virus hemagglutinin. Virology. 1986 Apr 30;150(2):479–490. doi: 10.1016/0042-6822(86)90312-0. [DOI] [PubMed] [Google Scholar]
  2. Becker P. B., Wu C. Cell-free system for assembly of transcriptionally repressed chromatin from Drosophila embryos. Mol Cell Biol. 1992 May;12(5):2241–2249. doi: 10.1128/mcb.12.5.2241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bell L. R., Maine E. M., Schedl P., Cline T. W. Sex-lethal, a Drosophila sex determination switch gene, exhibits sex-specific RNA splicing and sequence similarity to RNA binding proteins. Cell. 1988 Dec 23;55(6):1037–1046. doi: 10.1016/0092-8674(88)90248-6. [DOI] [PubMed] [Google Scholar]
  4. Boggs R. T., Gregor P., Idriss S., Belote J. M., McKeown M. Regulation of sexual differentiation in D. melanogaster via alternative splicing of RNA from the transformer gene. Cell. 1987 Aug 28;50(5):739–747. doi: 10.1016/0092-8674(87)90332-1. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Cavener D. R. Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates. Nucleic Acids Res. 1987 Feb 25;15(4):1353–1361. doi: 10.1093/nar/15.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chelsky D., Ralph R., Jonak G. Sequence requirements for synthetic peptide-mediated translocation to the nucleus. Mol Cell Biol. 1989 Jun;9(6):2487–2492. doi: 10.1128/mcb.9.6.2487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cline T. W. Autoregulatory functioning of a Drosophila gene product that establish es and maintains the sexually determined state. Genetics. 1984 Jun;107(2):231–277. doi: 10.1093/genetics/107.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Coward P., Nagai K., Chen D., Thomas H. D., Nagamine C. M., Lau Y. F. Polymorphism of a CAG trinucleotide repeat within Sry correlates with B6.YDom sex reversal. Nat Genet. 1994 Mar;6(3):245–250. doi: 10.1038/ng0394-245. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Giese K., Cox J., Grosschedl R. The HMG domain of lymphoid enhancer factor 1 bends DNA and facilitates assembly of functional nucleoprotein structures. Cell. 1992 Apr 3;69(1):185–195. doi: 10.1016/0092-8674(92)90129-z. [DOI] [PubMed] [Google Scholar]
  13. Goodfellow P. N., Lovell-Badge R. SRY and sex determination in mammals. Annu Rev Genet. 1993;27:71–92. doi: 10.1146/annurev.ge.27.120193.000443. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Granadino B., Penalva L. O., Sánchez L. The gene fl(2)d is needed for the sex-specific splicing of transformer pre-mRNA but not for double-sex pre-mRNA in Drosophila melanogaster. Mol Gen Genet. 1996 Nov 27;253(1-2):26–31. doi: 10.1007/s004380050292. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Gubbay J., Vivian N., Economou A., Jackson D., Goodfellow P., Lovell-Badge R. Inverted repeat structure of the Sry locus in mice. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7953–7957. doi: 10.1073/pnas.89.17.7953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hager J. H., Cline T. W. Induction of female Sex-lethal RNA splicing in male germ cells: implications for Drosophila germline sex determination. Development. 1997 Dec;124(24):5033–5048. doi: 10.1242/dev.124.24.5033. [DOI] [PubMed] [Google Scholar]
  19. Hilfiker A., Amrein H., Dübendorfer A., Schneiter R., Nöthiger R. The gene virilizer is required for female-specific splicing controlled by Sxl, the master gene for sexual development in Drosophila. Development. 1995 Dec;121(12):4017–4026. doi: 10.1242/dev.121.12.4017. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Koopman P., Gubbay J., Vivian N., Goodfellow P., Lovell-Badge R. Male development of chromosomally female mice transgenic for Sry. Nature. 1991 May 9;351(6322):117–121. doi: 10.1038/351117a0. [DOI] [PubMed] [Google Scholar]
  22. Lupas A. Coiled coils: new structures and new functions. Trends Biochem Sci. 1996 Oct;21(10):375–382. [PubMed] [Google Scholar]
  23. McKeown M., Belote J. M., Boggs R. T. Ectopic expression of the female transformer gene product leads to female differentiation of chromosomally male Drosophila. Cell. 1988 Jun 17;53(6):887–895. doi: 10.1016/s0092-8674(88)90369-8. [DOI] [PubMed] [Google Scholar]
  24. Nasrin N., Buggs C., Kong X. F., Carnazza J., Goebl M., Alexander-Bridges M. DNA-binding properties of the product of the testis-determining gene and a related protein. Nature. 1991 Nov 28;354(6351):317–320. doi: 10.1038/354317a0. [DOI] [PubMed] [Google Scholar]
  25. Nöthiger R., Jonglez M., Leuthold M., Meier-Gerschwiler P., Weber T. Sex determination in the germ line of Drosophila depends on genetic signals and inductive somatic factors. Development. 1989 Nov;107(3):505–518. doi: 10.1242/dev.107.3.505. [DOI] [PubMed] [Google Scholar]
  26. O'Hare K., Rubin G. M. Structures of P transposable elements and their sites of insertion and excision in the Drosophila melanogaster genome. Cell. 1983 Aug;34(1):25–35. doi: 10.1016/0092-8674(83)90133-2. [DOI] [PubMed] [Google Scholar]
  27. Oliver B., Perrimon N., Mahowald A. P. Genetic evidence that the sans fille locus is involved in Drosophila sex determination. Genetics. 1988 Sep;120(1):159–171. doi: 10.1093/genetics/120.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Penalva L. O., Sakamoto H., Navarro-Sabaté A., Sakashita E., Granadino B., Segarra C., Sánchez L. Regulation of the gene Sex-lethal: a comparative analysis of Drosophila melanogaster and Drosophila subobscura. Genetics. 1996 Dec;144(4):1653–1664. doi: 10.1093/genetics/144.4.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Perutz M. F. Glutamine repeats and neurodegenerative diseases: molecular aspects. Trends Biochem Sci. 1999 Feb;24(2):58–63. doi: 10.1016/s0968-0004(98)01350-4. [DOI] [PubMed] [Google Scholar]
  30. Perutz M. F., Staden R., Moens L., De Baere I. Polar zippers. Curr Biol. 1993 May 1;3(5):249–253. doi: 10.1016/0960-9822(93)90174-m. [DOI] [PubMed] [Google Scholar]
  31. Ptashne M., Gann A. Transcriptional activation by recruitment. Nature. 1997 Apr 10;386(6625):569–577. doi: 10.1038/386569a0. [DOI] [PubMed] [Google Scholar]
  32. Puig O., Gottschalk A., Fabrizio P., Séraphin B. Interaction of the U1 snRNP with nonconserved intronic sequences affects 5' splice site selection. Genes Dev. 1999 Mar 1;13(5):569–580. doi: 10.1101/gad.13.5.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Salz H. K., Cline T. W., Schedl P. Functional changes associated with structural alterations induced by mobilization of a P element inserted in the Sex-lethal gene of Drosophila. Genetics. 1987 Oct;117(2):221–231. doi: 10.1093/genetics/117.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Salz H. K., Maine E. M., Keyes L. N., Samuels M. E., Cline T. W., Schedl P. The Drosophila female-specific sex-determination gene, Sex-lethal, has stage-, tissue-, and sex-specific RNAs suggesting multiple modes of regulation. Genes Dev. 1989 May;3(5):708–719. doi: 10.1101/gad.3.5.708. [DOI] [PubMed] [Google Scholar]
  36. Salz H. K. The genetic analysis of snf: a Drosophila sex determination gene required for activation of Sex-lethal in both the germline and the soma. Genetics. 1992 Mar;130(3):547–554. doi: 10.1093/genetics/130.3.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schüpbach T. Normal female germ cell differentiation requires the female X chromosome to autosome ratio and expression of sex-lethal in Drosophila melanogaster. Genetics. 1985 Mar;109(3):529–548. doi: 10.1093/genetics/109.3.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Steinmann-Zwicky M., Schmid H., Nöthiger R. Cell-autonomous and inductive signals can determine the sex of the germ line of drosophila by regulating the gene Sxl. Cell. 1989 Apr 7;57(1):157–166. doi: 10.1016/0092-8674(89)90181-5. [DOI] [PubMed] [Google Scholar]
  40. Steinmann-Zwicky M. Sex determination in Drosophila: the X-chromosomal gene liz is required for Sxl activity. EMBO J. 1988 Dec 1;7(12):3889–3898. doi: 10.1002/j.1460-2075.1988.tb03275.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES