Skip to main content
PMC home page
Genetics logoLink to Genetics
. 2004 May;167(1):325–342. doi: 10.1534/genetics.167.1.325

An F1 genetic screen for maternal-effect mutations affecting embryonic pattern formation in Drosophila melanogaster.

Stefan Luschnig 1, Bernard Moussian 1, Jana Krauss 1, Isabelle Desjeux 1, Josip Perkovic 1, Christiane Nüsslein-Volhard 1
PMCID: PMC1470860  PMID: 15166158

Abstract

Large-scale screens for female-sterile mutations have revealed genes required maternally for establishment of the body axes in the Drosophila embryo. Although it is likely that the majority of components involved in axis formation have been identified by this approach, certain genes have escaped detection. This may be due to (1) incomplete saturation of the screens for female-sterile mutations and (2) genes with essential functions in zygotic development that mutate to lethality, precluding their identification as female-sterile mutations. To overcome these limitations, we performed a genetic mosaic screen aimed at identifying new maternal genes required for early embryonic patterning, including zygotically required ones. Using the Flp-FRT technique and a visible germline clone marker, we developed a system that allows efficient screening for maternal-effect phenotypes after only one generation of breeding, rather than after the three generations required for classic female-sterile screens. We identified 232 mutants showing various defects in embryonic pattern or morphogenesis. The mutants were ordered into 10 different phenotypic classes. A total of 174 mutants were assigned to 86 complementation groups with two alleles on average. Mutations in 45 complementation groups represent most previously known maternal genes, while 41 complementation groups represent new loci, including several involved in dorsoventral, anterior-posterior, and terminal patterning.

Full Text

The Full Text of this article is available as a PDF (680.3 KB).

Selected References

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

  1. Adams M. D., Celniker S. E., Holt R. A., Evans C. A., Gocayne J. D., Amanatides P. G., Scherer S. E., Li P. W., Hoskins R. A., Galle R. F. The genome sequence of Drosophila melanogaster. Science. 2000 Mar 24;287(5461):2185–2195. doi: 10.1126/science.287.5461.2185. [DOI] [PubMed] [Google Scholar]
  2. Ambrosio L., Mahowald A. P., Perrimon N. Requirement of the Drosophila raf homologue for torso function. Nature. 1989 Nov 16;342(6247):288–291. doi: 10.1038/342288a0. [DOI] [PubMed] [Google Scholar]
  3. Arrizabalaga G., Lehmann R. A selective screen reveals discrete functional domains in Drosophila Nanos. Genetics. 1999 Dec;153(4):1825–1838. doi: 10.1093/genetics/153.4.1825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ashburner M., Misra S., Roote J., Lewis S. E., Blazej R., Davis T., Doyle C., Galle R., George R., Harris N. An exploration of the sequence of a 2.9-Mb region of the genome of Drosophila melanogaster: the Adh region. Genetics. 1999 Sep;153(1):179–219. doi: 10.1093/genetics/153.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bellotto Manolo, Bopp Daniel, Senti Kirsten A., Burke Richard, Deak Peter, Maroy Peter, Dickson Barry, Basler Konrad, Hafen Ernst. Maternal-effect loci involved in Drosophila oogenesis and embryogenesis: P element-induced mutations on the third chromosome. Int J Dev Biol. 2002 Jan;46(1):149–157. [PubMed] [Google Scholar]
  6. Berger J., Suzuki T., Senti K. A., Stubbs J., Schaffner G., Dickson B. J. Genetic mapping with SNP markers in Drosophila. Nat Genet. 2001 Dec;29(4):475–481. doi: 10.1038/ng773. [DOI] [PubMed] [Google Scholar]
  7. Binari R., Perrimon N. Stripe-specific regulation of pair-rule genes by hopscotch, a putative Jak family tyrosine kinase in Drosophila. Genes Dev. 1994 Feb 1;8(3):300–312. doi: 10.1101/gad.8.3.300. [DOI] [PubMed] [Google Scholar]
  8. Casanova J., Furriols M., McCormick C. A., Struhl G. Similarities between trunk and spätzle, putative extracellular ligands specifying body pattern in Drosophila. Genes Dev. 1995 Oct 15;9(20):2539–2544. doi: 10.1101/gad.9.20.2539. [DOI] [PubMed] [Google Scholar]
  9. Chagnovich D., Lehmann R. Poly(A)-independent regulation of maternal hunchback translation in the Drosophila embryo. Proc Natl Acad Sci U S A. 2001 Sep 18;98(20):11359–11364. doi: 10.1073/pnas.201284398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Charatsi Iphigenie, Luschnig Stefan, Bartoszewski Slawomir, Nüsslein-Volhard Christiane, Moussian Bernard. Krapfen/dMyd88 is required for the establishment of dorsoventral pattern in the Drosophila embryo. Mech Dev. 2003 Feb;120(2):219–226. doi: 10.1016/s0925-4773(02)00410-0. [DOI] [PubMed] [Google Scholar]
  11. Chou T. B., Noll E., Perrimon N. Autosomal P[ovoD1] dominant female-sterile insertions in Drosophila and their use in generating germ-line chimeras. Development. 1993 Dec;119(4):1359–1369. doi: 10.1242/dev.119.4.1359. [DOI] [PubMed] [Google Scholar]
  12. Chou T. B., Perrimon N. The autosomal FLP-DFS technique for generating germline mosaics in Drosophila melanogaster. Genetics. 1996 Dec;144(4):1673–1679. doi: 10.1093/genetics/144.4.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chou T. B., Perrimon N. Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila. Genetics. 1992 Jul;131(3):643–653. doi: 10.1093/genetics/131.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Daubresse G., Deuring R., Moore L., Papoulas O., Zakrajsek I., Waldrip W. R., Scott M. P., Kennison J. A., Tamkun J. W. The Drosophila kismet gene is related to chromatin-remodeling factors and is required for both segmentation and segment identity. Development. 1999 Mar;126(6):1175–1187. doi: 10.1242/dev.126.6.1175. [DOI] [PubMed] [Google Scholar]
  15. Davis I., Girdham C. H., O'Farrell P. H. A nuclear GFP that marks nuclei in living Drosophila embryos; maternal supply overcomes a delay in the appearance of zygotic fluorescence. Dev Biol. 1995 Aug;170(2):726–729. doi: 10.1006/dbio.1995.1251. [DOI] [PubMed] [Google Scholar]
  16. Endow S. A., Chandra R., Komma D. J., Yamamoto A. H., Salmon E. D. Mutants of the Drosophila ncd microtubule motor protein cause centrosomal and spindle pole defects in mitosis. J Cell Sci. 1994 Apr;107(Pt 4):859–867. doi: 10.1242/jcs.107.4.859. [DOI] [PubMed] [Google Scholar]
  17. Erkner Alfrun, Roure Agnès, Charroux Bernard, Delaage Michèle, Holway Nicolas, Coré Nathalie, Vola Christine, Angelats Corinne, Pagès Françoise, Fasano Laurent. Grunge, related to human Atrophin-like proteins, has multiple functions in Drosophila development. Development. 2002 Mar;129(5):1119–1129. doi: 10.1242/dev.129.5.1119. [DOI] [PubMed] [Google Scholar]
  18. Furriols Marc, Casanova Jordi. In and out of Torso RTK signalling. EMBO J. 2003 May 1;22(9):1947–1952. doi: 10.1093/emboj/cdg224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gans M., Audit C., Masson M. Isolation and characterization of sex-linked female-sterile mutants in Drosophila melanogaster. Genetics. 1975 Dec;81(4):683–704. doi: 10.1093/genetics/81.4.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Garcia-Bellido A., Robbins L. G. Viability of Female Germ-Line Cells Homozygous for Zygotic Lethals in DROSOPHILA MELANOGASTER. Genetics. 1983 Feb;103(2):235–247. doi: 10.1093/genetics/103.2.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Goff D. J., Nilson L. A., Morisato D. Establishment of dorsal-ventral polarity of the Drosophila egg requires capicua action in ovarian follicle cells. Development. 2001 Nov;128(22):4553–4562. doi: 10.1242/dev.128.22.4553. [DOI] [PubMed] [Google Scholar]
  22. Golic K. G., Lindquist S. The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome. Cell. 1989 Nov 3;59(3):499–509. doi: 10.1016/0092-8674(89)90033-0. [DOI] [PubMed] [Google Scholar]
  23. Golic K. G. Site-specific recombination between homologous chromosomes in Drosophila. Science. 1991 May 17;252(5008):958–961. doi: 10.1126/science.2035025. [DOI] [PubMed] [Google Scholar]
  24. Grether M. E., Abrams J. M., Agapite J., White K., Steller H. The head involution defective gene of Drosophila melanogaster functions in programmed cell death. Genes Dev. 1995 Jul 15;9(14):1694–1708. doi: 10.1101/gad.9.14.1694. [DOI] [PubMed] [Google Scholar]
  25. Grosshans J., Schnorrer F., Nüsslein-Volhard C. Oligomerisation of Tube and Pelle leads to nuclear localisation of dorsal. Mech Dev. 1999 Mar;81(1-2):127–138. doi: 10.1016/s0925-4773(98)00236-6. [DOI] [PubMed] [Google Scholar]
  26. Guichet A., Copeland J. W., Erdélyi M., Hlousek D., Závorszky P., Ho J., Brown S., Percival-Smith A., Krause H. M., Ephrussi A. The nuclear receptor homologue Ftz-F1 and the homeodomain protein Ftz are mutually dependent cofactors. Nature. 1997 Feb 6;385(6616):548–552. doi: 10.1038/385548a0. [DOI] [PubMed] [Google Scholar]
  27. Hou X. S., Chou T. B., Melnick M. B., Perrimon N. The torso receptor tyrosine kinase can activate Raf in a Ras-independent pathway. Cell. 1995 Apr 7;81(1):63–71. doi: 10.1016/0092-8674(95)90371-2. [DOI] [PubMed] [Google Scholar]
  28. Jiménez Gerardo, González-Reyes Acaimo, Casanova Jordi. Cell surface proteins Nasrat and Polehole stabilize the Torso-like extracellular determinant in Drosophila oogenesis. Genes Dev. 2002 Apr 15;16(8):913–918. doi: 10.1101/gad.223902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Luschnig S., Krauss J., Bohmann K., Desjeux I., Nüsslein-Volhard C. The Drosophila SHC adaptor protein is required for signaling by a subset of receptor tyrosine kinases. Mol Cell. 2000 Feb;5(2):231–241. doi: 10.1016/s1097-2765(00)80419-0. [DOI] [PubMed] [Google Scholar]
  30. Mlodzik M., Gehring W. J. Expression of the caudal gene in the germ line of Drosophila: formation of an RNA and protein gradient during early embryogenesis. Cell. 1987 Feb 13;48(3):465–478. doi: 10.1016/0092-8674(87)90197-8. [DOI] [PubMed] [Google Scholar]
  31. Moore L. A., Broihier H. T., Van Doren M., Lunsford L. B., Lehmann R. Identification of genes controlling germ cell migration and embryonic gonad formation in Drosophila. Development. 1998 Feb;125(4):667–678. doi: 10.1242/dev.125.4.667. [DOI] [PubMed] [Google Scholar]
  32. Mével-Ninio M., Fouilloux E., Guénal I., Vincent A. The three dominant female-sterile mutations of the Drosophila ovo gene are point mutations that create new translation-initiator AUG codons. Development. 1996 Dec;122(12):4131–4138. doi: 10.1242/dev.122.12.4131. [DOI] [PubMed] [Google Scholar]
  33. Nairz Knud, Stocker Hugo, Schindelholz Benno, Hafen Ernst. High-resolution SNP mapping by denaturing HPLC. Proc Natl Acad Sci U S A. 2002 Jul 29;99(16):10575–10580. doi: 10.1073/pnas.162136299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Parkhurst S. M., Ish-Horowicz D. wimp, a dominant maternal-effect mutation, reduces transcription of a specific subset of segmentation genes in Drosophila. Genes Dev. 1991 Mar;5(3):341–357. doi: 10.1101/gad.5.3.341. [DOI] [PubMed] [Google Scholar]
  36. Perrimon N., Engstrom L., Mahowald A. P. The effects of zygotic lethal mutations on female germ-line functions in Drosophila. Dev Biol. 1984 Oct;105(2):404–414. doi: 10.1016/0012-1606(84)90297-5. [DOI] [PubMed] [Google Scholar]
  37. Perrimon N., Lanjuin A., Arnold C., Noll E. Zygotic lethal mutations with maternal effect phenotypes in Drosophila melanogaster. II. Loci on the second and third chromosomes identified by P-element-induced mutations. Genetics. 1996 Dec;144(4):1681–1692. doi: 10.1093/genetics/144.4.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Perrimon N., Mohler D., Engstrom L., Mahowald A. P. X-linked female-sterile loci in Drosophila melanogaster. Genetics. 1986 Jul;113(3):695–712. doi: 10.1093/genetics/113.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Raabe T., Riesgo-Escovar J., Liu X., Bausenwein B. S., Deak P., Maröy P., Hafen E. DOS, a novel pleckstrin homology domain-containing protein required for signal transduction between sevenless and Ras1 in Drosophila. Cell. 1996 Jun 14;85(6):911–920. doi: 10.1016/s0092-8674(00)81274-x. [DOI] [PubMed] [Google Scholar]
  40. Rittenhouse K. R., Berg C. A. Mutations in the Drosophila gene bullwinkle cause the formation of abnormal eggshell structures and bicaudal embryos. Development. 1995 Sep;121(9):3023–3033. doi: 10.1242/dev.121.9.3023. [DOI] [PubMed] [Google Scholar]
  41. Roch Fernando, Jiménez Gerardo, Casanova Jordi. EGFR signalling inhibits Capicua-dependent repression during specification of Drosophila wing veins. Development. 2002 Feb;129(4):993–1002. doi: 10.1242/dev.129.4.993. [DOI] [PubMed] [Google Scholar]
  42. Rubin G. M., Yandell M. D., Wortman J. R., Gabor Miklos G. L., Nelson C. R., Hariharan I. K., Fortini M. E., Li P. W., Apweiler R., Fleischmann W. Comparative genomics of the eukaryotes. Science. 2000 Mar 24;287(5461):2204–2215. doi: 10.1126/science.287.5461.2204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Schnorrer Frank, Luschnig Stefan, Koch Iris, Nüsslein-Volhard Christiane. Gamma-tubulin37C and gamma-tubulin ring complex protein 75 are essential for bicoid RNA localization during drosophila oogenesis. Dev Cell. 2002 Nov;3(5):685–696. doi: 10.1016/s1534-5807(02)00301-5. [DOI] [PubMed] [Google Scholar]
  44. Schupbach T., Wieschaus E. Germline autonomy of maternal-effect mutations altering the embryonic body pattern of Drosophila. Dev Biol. 1986 Feb;113(2):443–448. doi: 10.1016/0012-1606(86)90179-x. [DOI] [PubMed] [Google Scholar]
  45. Schüpbach T., Wieschaus E. Female sterile mutations on the second chromosome of Drosophila melanogaster. I. Maternal effect mutations. Genetics. 1989 Jan;121(1):101–117. doi: 10.1093/genetics/121.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Simon M. A., Bowtell D. D., Dodson G. S., Laverty T. R., Rubin G. M. Ras1 and a putative guanine nucleotide exchange factor perform crucial steps in signaling by the sevenless protein tyrosine kinase. Cell. 1991 Nov 15;67(4):701–716. doi: 10.1016/0092-8674(91)90065-7. [DOI] [PubMed] [Google Scholar]
  47. Sonoda J., Wharton R. P. Drosophila Brain Tumor is a translational repressor. Genes Dev. 2001 Mar 15;15(6):762–773. doi: 10.1101/gad.870801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. St Johnston D., Nüsslein-Volhard C. The origin of pattern and polarity in the Drosophila embryo. Cell. 1992 Jan 24;68(2):201–219. doi: 10.1016/0092-8674(92)90466-p. [DOI] [PubMed] [Google Scholar]
  49. St Johnston Daniel. The art and design of genetic screens: Drosophila melanogaster. Nat Rev Genet. 2002 Mar;3(3):176–188. doi: 10.1038/nrg751. [DOI] [PubMed] [Google Scholar]
  50. Stern C. Somatic Crossing over and Segregation in Drosophila Melanogaster. Genetics. 1936 Nov;21(6):625–730. doi: 10.1093/genetics/21.6.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Struhl G. A gene product required for correct initiation of segmental determination in Drosophila. Nature. 1981 Sep 3;293(5827):36–41. doi: 10.1038/293036a0. [DOI] [PubMed] [Google Scholar]
  52. Struhl G., Basler K. Organizing activity of wingless protein in Drosophila. Cell. 1993 Feb 26;72(4):527–540. doi: 10.1016/0092-8674(93)90072-x. [DOI] [PubMed] [Google Scholar]
  53. Su M. A., Wisotzkey R. G., Newfeld S. J. A screen for modifiers of decapentaplegic mutant phenotypes identifies lilliputian, the only member of the Fragile-X/Burkitt's Lymphoma family of transcription factors in Drosophila melanogaster. Genetics. 2001 Feb;157(2):717–725. doi: 10.1093/genetics/157.2.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Tang A. H., Neufeld T. P., Rubin G. M., Müller H. A. Transcriptional regulation of cytoskeletal functions and segmentation by a novel maternal pair-rule gene, lilliputian. Development. 2001 Mar;128(5):801–813. doi: 10.1242/dev.128.5.801. [DOI] [PubMed] [Google Scholar]
  55. Therrien M., Chang H. C., Solomon N. M., Karim F. D., Wassarman D. A., Rubin G. M. KSR, a novel protein kinase required for RAS signal transduction. Cell. 1995 Dec 15;83(6):879–888. doi: 10.1016/0092-8674(95)90204-x. [DOI] [PubMed] [Google Scholar]
  56. Treisman J. E., Luk A., Rubin G. M., Heberlein U. eyelid antagonizes wingless signaling during Drosophila development and has homology to the Bright family of DNA-binding proteins. Genes Dev. 1997 Aug 1;11(15):1949–1962. doi: 10.1101/gad.11.15.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Ueno S., Kondoh K., Kotani Y., Komure O., Kuno S., Kawai J., Hazama F., Sano A. Somatic mosaicism of CAG repeat in dentatorubral-pallidoluysian atrophy (DRPLA). Hum Mol Genet. 1995 Apr;4(4):663–666. doi: 10.1093/hmg/4.4.663. [DOI] [PubMed] [Google Scholar]
  58. Vázquez M., Moore L., Kennison J. A. The trithorax group gene osa encodes an ARID-domain protein that genetically interacts with the brahma chromatin-remodeling factor to regulate transcription. Development. 1999 Feb;126(4):733–742. doi: 10.1242/dev.126.4.733. [DOI] [PubMed] [Google Scholar]
  59. Wittwer F., van der Straten A., Keleman K., Dickson B. J., Hafen E. Lilliputian: an AF4/FMR2-related protein that controls cell identity and cell growth. Development. 2001 Mar;128(5):791–800. doi: 10.1242/dev.128.5.791. [DOI] [PubMed] [Google Scholar]
  60. Wodarz A., Ramrath A., Grimm A., Knust E. Drosophila atypical protein kinase C associates with Bazooka and controls polarity of epithelia and neuroblasts. J Cell Biol. 2000 Sep 18;150(6):1361–1374. doi: 10.1083/jcb.150.6.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Xie T., Spradling A. C. decapentaplegic is essential for the maintenance and division of germline stem cells in the Drosophila ovary. Cell. 1998 Jul 24;94(2):251–260. doi: 10.1016/s0092-8674(00)81424-5. [DOI] [PubMed] [Google Scholar]
  62. Xu T., Rubin G. M. Analysis of genetic mosaics in developing and adult Drosophila tissues. Development. 1993 Apr;117(4):1223–1237. doi: 10.1242/dev.117.4.1223. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES