Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1996 Feb 1;15(3):640–649.

Asymmetric localization of Drosophila pair-rule transcripts from displaced nuclei: evidence for directional nuclear export.

H Francis-Lang 1, I Davis 1, D Ish-Horowicz 1
PMCID: PMC449982  PMID: 8599947

Abstract

Drosophila pair-rule transcripts accumulate exclusively apical of the layer of peripheral nuclei in syncytial blastoderm stage embryos. Here, we use aneuploid embryos to test zygotic gene requirements for pair-rule transcript localization. As apical localization is maintained in all genotypes tested, the required components must be maternally encoded. In aneuploid embryos with multiple layers or cortical nuclei, pair-rule transcripts lie apical of both superficial and internalized nuclei. In the latter case, the transcripts are 'pseudo-apical', i.e. apical of the nuclei from which they derive, but basal of superficial nuclei. We show that internalized nuclei maintain their apico-basal nuclear orientation, and that they lack the apical cytoskeletal assemblies which lie adjacent to superficial nuclei. These results support a mechanism of localizing pair-rule transcripts by directional (vectorial) nuclear export.

Full text

PDF
640

Images in this article

641Image
on p.641
642Image
on p.642
643Image
on p.643
644Image
on p.644
645Image
on p.645
645Image
on p.645

Selected References

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

  1. Bassell G. J., Powers C. M., Taneja K. L., Singer R. H. Single mRNAs visualized by ultrastructural in situ hybridization are principally localized at actin filament intersections in fibroblasts. J Cell Biol. 1994 Aug;126(4):863–876. doi: 10.1083/jcb.126.4.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berleth T., Burri M., Thoma G., Bopp D., Richstein S., Frigerio G., Noll M., Nüsslein-Volhard C. The role of localization of bicoid RNA in organizing the anterior pattern of the Drosophila embryo. EMBO J. 1988 Jun;7(6):1749–1756. doi: 10.1002/j.1460-2075.1988.tb03004.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blobel G. Gene gating: a hypothesis. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8527–8529. doi: 10.1073/pnas.82.24.8527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark I., Giniger E., Ruohola-Baker H., Jan L. Y., Jan Y. N. Transient posterior localization of a kinesin fusion protein reflects anteroposterior polarity of the Drosophila oocyte. Curr Biol. 1994 Apr 1;4(4):289–300. doi: 10.1016/s0960-9822(00)00068-3. [DOI] [PubMed] [Google Scholar]
  5. Comings D. E., Drets M. E. Mechanisms of chromosome banding. IX. Are variations in DNA base composition adequate to account for quinacrine, Hoechst 33258 and daunomycin banding? Chromosoma. 1976 Jul 8;56(3):199–211. doi: 10.1007/BF00293185. [DOI] [PubMed] [Google Scholar]
  6. Davis I., Francis-Lang H., Ish-Horowicz D. Mechanisms of intracellular transcript localization and export in early Drosophila embryos. Cold Spring Harb Symp Quant Biol. 1993;58:793–798. doi: 10.1101/sqb.1993.058.01.086. [DOI] [PubMed] [Google Scholar]
  7. Davis I., Ish-Horowicz D. Apical localization of pair-rule transcripts requires 3' sequences and limits protein diffusion in the Drosophila blastoderm embryo. Cell. 1991 Nov 29;67(5):927–940. doi: 10.1016/0092-8674(91)90366-7. [DOI] [PubMed] [Google Scholar]
  8. Edgar B. A., Odell G. M., Schubiger G. Cytoarchitecture and the patterning of fushi tarazu expression in the Drosophila blastoderm. Genes Dev. 1987 Dec;1(10):1226–1237. doi: 10.1101/gad.1.10.1226. [DOI] [PubMed] [Google Scholar]
  9. Edgar B. A., Weir M. P., Schubiger G., Kornberg T. Repression and turnover pattern fushi tarazu RNA in the early Drosophila embryo. Cell. 1986 Dec 5;47(5):747–754. doi: 10.1016/0092-8674(86)90517-9. [DOI] [PubMed] [Google Scholar]
  10. Ephrussi A., Dickinson L. K., Lehmann R. Oskar organizes the germ plasm and directs localization of the posterior determinant nanos. Cell. 1991 Jul 12;66(1):37–50. doi: 10.1016/0092-8674(91)90137-n. [DOI] [PubMed] [Google Scholar]
  11. Ephrussi A., Lehmann R. Induction of germ cell formation by oskar. Nature. 1992 Jul 30;358(6385):387–392. doi: 10.1038/358387a0. [DOI] [PubMed] [Google Scholar]
  12. Ferrandon D., Elphick L., Nüsslein-Volhard C., St Johnston D. Staufen protein associates with the 3'UTR of bicoid mRNA to form particles that move in a microtubule-dependent manner. Cell. 1994 Dec 30;79(7):1221–1232. doi: 10.1016/0092-8674(94)90013-2. [DOI] [PubMed] [Google Scholar]
  13. Gavis E. R., Lehmann R. Localization of nanos RNA controls embryonic polarity. Cell. 1992 Oct 16;71(2):301–313. doi: 10.1016/0092-8674(92)90358-j. [DOI] [PubMed] [Google Scholar]
  14. González-Reyes A., Elliott H., St Johnston D. Polarization of both major body axes in Drosophila by gurken-torpedo signalling. Nature. 1995 Jun 22;375(6533):654–658. doi: 10.1038/375654a0. [DOI] [PubMed] [Google Scholar]
  15. González-Reyes A., St Johnston D. Role of oocyte position in establishment of anterior-posterior polarity in Drosophila. Science. 1994 Oct 28;266(5185):639–642. doi: 10.1126/science.7939717. [DOI] [PubMed] [Google Scholar]
  16. Hafen E., Kuroiwa A., Gehring W. J. Spatial distribution of transcripts from the segmentation gene fushi tarazu during Drosophila embryonic development. Cell. 1984 Jul;37(3):833–841. doi: 10.1016/0092-8674(84)90418-5. [DOI] [PubMed] [Google Scholar]
  17. Hiraoka Y., Agard D. A., Sedat J. W. Temporal and spatial coordination of chromosome movement, spindle formation, and nuclear envelope breakdown during prometaphase in Drosophila melanogaster embryos. J Cell Biol. 1990 Dec;111(6 Pt 2):2815–2828. doi: 10.1083/jcb.111.6.2815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jongens T. A., Ackerman L. D., Swedlow J. R., Jan L. Y., Jan Y. N. Germ cell-less encodes a cell type-specific nuclear pore-associated protein and functions early in the germ-cell specification pathway of Drosophila. Genes Dev. 1994 Sep 15;8(18):2123–2136. doi: 10.1101/gad.8.18.2123. [DOI] [PubMed] [Google Scholar]
  19. Karr T. L., Alberts B. M. Organization of the cytoskeleton in early Drosophila embryos. J Cell Biol. 1986 Apr;102(4):1494–1509. doi: 10.1083/jcb.102.4.1494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kislauskis E. H., Li Z., Singer R. H., Taneja K. L. Isoform-specific 3'-untranslated sequences sort alpha-cardiac and beta-cytoplasmic actin messenger RNAs to different cytoplasmic compartments. J Cell Biol. 1993 Oct;123(1):165–172. doi: 10.1083/jcb.123.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kislauskis E. H., Zhu X., Singer R. H. Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype. J Cell Biol. 1994 Oct;127(2):441–451. doi: 10.1083/jcb.127.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lane M. E., Kalderon D. RNA localization along the anteroposterior axis of the Drosophila oocyte requires PKA-mediated signal transduction to direct normal microtubule organization. Genes Dev. 1994 Dec 15;8(24):2986–2995. doi: 10.1101/gad.8.24.2986. [DOI] [PubMed] [Google Scholar]
  23. Litman P., Barg J., Ginzburg I. Microtubules are involved in the localization of tau mRNA in primary neuronal cell cultures. Neuron. 1994 Dec;13(6):1463–1474. doi: 10.1016/0896-6273(94)90432-4. [DOI] [PubMed] [Google Scholar]
  24. Macdonald P. M., Kerr K., Smith J. L., Leask A. RNA regulatory element BLE1 directs the early steps of bicoid mRNA localization. Development. 1993 Aug;118(4):1233–1243. doi: 10.1242/dev.118.4.1233. [DOI] [PubMed] [Google Scholar]
  25. Merrill P. T., Sweeton D., Wieschaus E. Requirements for autosomal gene activity during precellular stages of Drosophila melanogaster. Development. 1988 Nov;104(3):495–509. doi: 10.1242/dev.104.3.495. [DOI] [PubMed] [Google Scholar]
  26. Pelham H. R., Munro S. Sorting of membrane proteins in the secretory pathway. Cell. 1993 Nov 19;75(4):603–605. doi: 10.1016/0092-8674(93)90479-A. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pokrywka N. J., Stephenson E. C. Localized RNAs are enriched in cytoskeletal extracts of Drosophila oocytes. Dev Biol. 1994 Nov;166(1):210–219. doi: 10.1006/dbio.1994.1308. [DOI] [PubMed] [Google Scholar]
  28. Pokrywka N. J., Stephenson E. C. Microtubules are a general component of mRNA localization systems in Drosophila oocytes. Dev Biol. 1995 Jan;167(1):363–370. doi: 10.1006/dbio.1995.1030. [DOI] [PubMed] [Google Scholar]
  29. Pokrywka N. J., Stephenson E. C. Microtubules mediate the localization of bicoid RNA during Drosophila oogenesis. Development. 1991 Sep;113(1):55–66. doi: 10.1242/dev.113.1.55. [DOI] [PubMed] [Google Scholar]
  30. Pondel M. D., King M. L. Localized maternal mRNA related to transforming growth factor beta mRNA is concentrated in a cytokeratin-enriched fraction from Xenopus oocytes. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7612–7616. doi: 10.1073/pnas.85.20.7612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rosbash M., Singer R. H. RNA travel: tracks from DNA to cytoplasm. Cell. 1993 Nov 5;75(3):399–401. doi: 10.1016/0092-8674(93)90373-x. [DOI] [PubMed] [Google Scholar]
  32. Saitoh Y., Laemmli U. K. Metaphase chromosome structure: bands arise from a differential folding path of the highly AT-rich scaffold. Cell. 1994 Feb 25;76(4):609–622. doi: 10.1016/0092-8674(94)90502-9. [DOI] [PubMed] [Google Scholar]
  33. Schejter E. D., Wieschaus E. Functional elements of the cytoskeleton in the early Drosophila embryo. Annu Rev Cell Biol. 1993;9:67–99. doi: 10.1146/annurev.cb.09.110193.000435. [DOI] [PubMed] [Google Scholar]
  34. Schejter E. D., Wieschaus E. bottleneck acts as a regulator of the microfilament network governing cellularization of the Drosophila embryo. Cell. 1993 Oct 22;75(2):373–385. doi: 10.1016/0092-8674(93)80078-s. [DOI] [PubMed] [Google Scholar]
  35. Schweisguth F., Lepesant J. A., Vincent A. The serendipity alpha gene encodes a membrane-associated protein required for the cellularization of the Drosophila embryo. Genes Dev. 1990 Jun;4(6):922–931. doi: 10.1101/gad.4.6.922. [DOI] [PubMed] [Google Scholar]
  36. Shermoen A. W., O'Farrell P. H. Progression of the cell cycle through mitosis leads to abortion of nascent transcripts. Cell. 1991 Oct 18;67(2):303–310. doi: 10.1016/0092-8674(91)90182-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Simpson L., Wieschaus E. Zygotic activity of the nullo locus is required to stabilize the actin-myosin network during cellularization in Drosophila. Development. 1990 Nov;110(3):851–863. doi: 10.1242/dev.110.3.851. [DOI] [PubMed] [Google Scholar]
  38. Singer R. H. The cytoskeleton and mRNA localization. Curr Opin Cell Biol. 1992 Feb;4(1):15–19. doi: 10.1016/0955-0674(92)90053-f. [DOI] [PubMed] [Google Scholar]
  39. St Johnston D., Driever W., Berleth T., Richstein S., Nüsslein-Volhard C. Multiple steps in the localization of bicoid RNA to the anterior pole of the Drosophila oocyte. Development. 1989;107 (Suppl):13–19. doi: 10.1242/dev.107.Supplement.13. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. St Johnston D. The intracellular localization of messenger RNAs. Cell. 1995 Apr 21;81(2):161–170. doi: 10.1016/0092-8674(95)90324-0. [DOI] [PubMed] [Google Scholar]
  42. Stafstrom J. P., Staehelin L. A. Dynamics of the nuclear envelope and of nuclear pore complexes during mitosis in the Drosophila embryo. Eur J Cell Biol. 1984 May;34(1):179–189. [PubMed] [Google Scholar]
  43. Sundell C. L., Singer R. H. Requirement of microfilaments in sorting of actin messenger RNA. Science. 1991 Sep 13;253(5025):1275–1277. doi: 10.1126/science.1891715. [DOI] [PubMed] [Google Scholar]
  44. Tautz D., Pfeifle C. A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma. 1989 Aug;98(2):81–85. doi: 10.1007/BF00291041. [DOI] [PubMed] [Google Scholar]
  45. Theurkauf W. E., Alberts B. M., Jan Y. N., Jongens T. A. A central role for microtubules in the differentiation of Drosophila oocytes. Development. 1993 Aug;118(4):1169–1180. doi: 10.1242/dev.118.4.1169. [DOI] [PubMed] [Google Scholar]
  46. Theurkauf W. E. Behavior of structurally divergent alpha-tubulin isotypes during Drosophila embryogenesis: evidence for post-translational regulation of isotype abundance. Dev Biol. 1992 Nov;154(1):205–217. doi: 10.1016/0012-1606(92)90060-t. [DOI] [PubMed] [Google Scholar]
  47. Theurkauf W. E. Premature microtubule-dependent cytoplasmic streaming in cappuccino and spire mutant oocytes. Science. 1994 Sep 30;265(5181):2093–2096. doi: 10.1126/science.8091233. [DOI] [PubMed] [Google Scholar]
  48. Vavra S. H., Carroll S. B. The zygotic control of Drosophila pair-rule gene expression. I. A search for new pair-rule regulatory loci. Development. 1989 Nov;107(3):663–672. doi: 10.1242/dev.107.3.663. [DOI] [PubMed] [Google Scholar]
  49. Vincent J. P., O'Farrell P. H. The state of engrailed expression is not clonally transmitted during early Drosophila development. Cell. 1992 Mar 6;68(5):923–931. doi: 10.1016/0092-8674(92)90035-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wang S., Hazelrigg T. Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis. Nature. 1994 Jun 2;369(6479):400–403. doi: 10.1038/369400a0. [DOI] [PubMed] [Google Scholar]
  51. Whitfield W. G., Millar S. E., Saumweber H., Frasch M., Glover D. M. Cloning of a gene encoding an antigen associated with the centrosome in Drosophila. J Cell Sci. 1988 Apr;89(Pt 4):467–480. doi: 10.1242/jcs.89.4.467. [DOI] [PubMed] [Google Scholar]
  52. Wieschaus E., Sweeton D. Requirements for X-linked zygotic gene activity during cellularization of early Drosophila embryos. Development. 1988 Nov;104(3):483–493. doi: 10.1242/dev.104.3.483. [DOI] [PubMed] [Google Scholar]
  53. Wilhelm J. E., Vale R. D. RNA on the move: the mRNA localization pathway. J Cell Biol. 1993 Oct;123(2):269–274. doi: 10.1083/jcb.123.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yisraeli J. K., Sokol S., Melton D. A. A two-step model for the localization of maternal mRNA in Xenopus oocytes: involvement of microtubules and microfilaments in the translocation and anchoring of Vg1 mRNA. Development. 1990 Feb;108(2):289–298. doi: 10.1242/dev.108.2.289. [DOI] [PubMed] [Google Scholar]
  55. Zachar Z., Kramer J., Mims I. P., Bingham P. M. Evidence for channeled diffusion of pre-mRNAs during nuclear RNA transport in metazoans. J Cell Biol. 1993 May;121(4):729–742. doi: 10.1083/jcb.121.4.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. von Heijne G. Protein targeting signals. Curr Opin Cell Biol. 1990 Aug;2(4):604–608. doi: 10.1016/0955-0674(90)90100-s. [DOI] [PubMed] [Google Scholar]

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

RESOURCES