Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Jun 1;129(5):1205–1216. doi: 10.1083/jcb.129.5.1205

Structural interaction between the nuclear pore complex and a specific translocating RNP particle

PMCID: PMC2120458  PMID: 7775568

Abstract

The transport of Balbiani ring (BR) premessenger RNP particles in the larval salivary gland cells of the dipteran Chironomus tentans can be followed using electron microscopy. A BR RNP particle consists of an RNP ribbon bent into a ringlike structure. Upon translocation through the nuclear pore complex (NPC), the ribbon is straightened and enters the central channel of the NPC with the 5' end of the transcript in the lead. The translocating ribbon is likely to interact with the central channel but, in addition, the remaining portion of the ribbon ring makes contact with the periphery of the NPC. To determine the nature of this latter interaction, we have now studied the connections between the RNP particle and the border of the NPC during different stages of translocation using electron microscope tomography. It was observed that the 3' terminal domain of the ribbon always touches the nuclear ring of the NPC, but the precise area of contact is variable. Sometimes also a region on the opposite side of the ribbon ring reaches the nuclear ring. The pattern of contacts could be correlated to the stage of translocation, and it was concluded that the particle-nuclear ring interactions reflect a rotation of the ribbon ring in front of the central channel, the rotation being secondary to the successive translocation of the ribbon through the channel. The particle's mode of interaction with the NPC suggests that the initial contact between the 5' end domain of the ribbon and the entrance to the central channel is probably crucial to accomplish the ordered translocation of the premessenger RNP particle through the NPC.

Full Text

The Full Text of this article is available as a PDF (7.5 MB).

Selected References

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

  1. Akey C. W., Goldfarb D. S. Protein import through the nuclear pore complex is a multistep process. J Cell Biol. 1989 Sep;109(3):971–982. doi: 10.1083/jcb.109.3.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Akey C. W., Radermacher M. Architecture of the Xenopus nuclear pore complex revealed by three-dimensional cryo-electron microscopy. J Cell Biol. 1993 Jul;122(1):1–19. doi: 10.1083/jcb.122.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andersson K., Björkroth B., Daneholt B. The in situ structure of the active 75 S RNA genes in Balbiani rings of Chironomus tentans. Exp Cell Res. 1980 Dec;130(2):313–326. doi: 10.1016/0014-4827(80)90008-7. [DOI] [PubMed] [Google Scholar]
  4. Bataillé N., Helser T., Fried H. M. Cytoplasmic transport of ribosomal subunits microinjected into the Xenopus laevis oocyte nucleus: a generalized, facilitated process. J Cell Biol. 1990 Oct;111(4):1571–1582. doi: 10.1083/jcb.111.4.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cordes V. C., Reidenbach S., Köhler A., Stuurman N., van Driel R., Franke W. W. Intranuclear filaments containing a nuclear pore complex protein. J Cell Biol. 1993 Dec;123(6 Pt 1):1333–1344. doi: 10.1083/jcb.123.6.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dargemont C., Kühn L. C. Export of mRNA from microinjected nuclei of Xenopus laevis oocytes. J Cell Biol. 1992 Jul;118(1):1–9. doi: 10.1083/jcb.118.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dingwall C., Laskey R. The nuclear membrane. Science. 1992 Nov 6;258(5084):942–947. doi: 10.1126/science.1439805. [DOI] [PubMed] [Google Scholar]
  8. Dreyfuss G., Matunis M. J., Piñol-Roma S., Burd C. G. hnRNP proteins and the biogenesis of mRNA. Annu Rev Biochem. 1993;62:289–321. doi: 10.1146/annurev.bi.62.070193.001445. [DOI] [PubMed] [Google Scholar]
  9. Dworetzky S. I., Feldherr C. M. Translocation of RNA-coated gold particles through the nuclear pores of oocytes. J Cell Biol. 1988 Mar;106(3):575–584. doi: 10.1083/jcb.106.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Featherstone C., Darby M. K., Gerace L. A monoclonal antibody against the nuclear pore complex inhibits nucleocytoplasmic transport of protein and RNA in vivo. J Cell Biol. 1988 Oct;107(4):1289–1297. doi: 10.1083/jcb.107.4.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feldherr C. M. The uptake of endogenous proteins by oocyte nuclei. Exp Cell Res. 1975 Jul;93(2):411–419. doi: 10.1016/0014-4827(75)90467-x. [DOI] [PubMed] [Google Scholar]
  12. Frank J., Verschoor A., Boublik M. Computer averaging of electron micrographs of 40S ribosomal subunits. Science. 1981 Dec 18;214(4527):1353–1355. doi: 10.1126/science.7313694. [DOI] [PubMed] [Google Scholar]
  13. Gerace L. Molecular trafficking across the nuclear pore complex. Curr Opin Cell Biol. 1992 Aug;4(4):637–645. doi: 10.1016/0955-0674(92)90083-o. [DOI] [PubMed] [Google Scholar]
  14. Goldberg M. W., Allen T. D. High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores. J Cell Biol. 1992 Dec;119(6):1429–1440. doi: 10.1083/jcb.119.6.1429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Guddat U., Bakken A. H., Pieler T. Protein-mediated nuclear export of RNA: 5S rRNA containing small RNPs in xenopus oocytes. Cell. 1990 Feb 23;60(4):619–628. doi: 10.1016/0092-8674(90)90665-2. [DOI] [PubMed] [Google Scholar]
  16. Hamm J., Mattaj I. W. Monomethylated cap structures facilitate RNA export from the nucleus. Cell. 1990 Oct 5;63(1):109–118. doi: 10.1016/0092-8674(90)90292-m. [DOI] [PubMed] [Google Scholar]
  17. Hinshaw J. E., Carragher B. O., Milligan R. A. Architecture and design of the nuclear pore complex. Cell. 1992 Jun 26;69(7):1133–1141. doi: 10.1016/0092-8674(92)90635-p. [DOI] [PubMed] [Google Scholar]
  18. Izaurralde E., Mattaj I. W. Transport of RNA between nucleus and cytoplasm. Semin Cell Biol. 1992 Aug;3(4):279–288. doi: 10.1016/1043-4682(92)90029-u. [DOI] [PubMed] [Google Scholar]
  19. Izaurralde E., Stepinski J., Darzynkiewicz E., Mattaj I. W. A cap binding protein that may mediate nuclear export of RNA polymerase II-transcribed RNAs. J Cell Biol. 1992 Sep;118(6):1287–1295. doi: 10.1083/jcb.118.6.1287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jarmolowski A., Boelens W. C., Izaurralde E., Mattaj I. W. Nuclear export of different classes of RNA is mediated by specific factors. J Cell Biol. 1994 Mar;124(5):627–635. doi: 10.1083/jcb.124.5.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jarnik M., Aebi U. Toward a more complete 3-D structure of the nuclear pore complex. J Struct Biol. 1991 Dec;107(3):291–308. doi: 10.1016/1047-8477(91)90054-z. [DOI] [PubMed] [Google Scholar]
  22. Khanna-Gupta A., Ware V. C. Nucleocytoplasmic transport of ribosomes in a eukaryotic system: is there a facilitated transport process? Proc Natl Acad Sci U S A. 1989 Mar;86(6):1791–1795. doi: 10.1073/pnas.86.6.1791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kraemer D., Wozniak R. W., Blobel G., Radu A. The human CAN protein, a putative oncogene product associated with myeloid leukemogenesis, is a nuclear pore complex protein that faces the cytoplasm. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1519–1523. doi: 10.1073/pnas.91.4.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lang I., Scholz M., Peters R. Molecular mobility and nucleocytoplasmic flux in hepatoma cells. J Cell Biol. 1986 Apr;102(4):1183–1190. doi: 10.1083/jcb.102.4.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maul G. G. The nuclear and the cytoplasmic pore complex: structure, dynamics, distribution, and evolution. Int Rev Cytol Suppl. 1977;(6):75–186. [PubMed] [Google Scholar]
  26. Mehlin H., Daneholt B., Skoglund U. Translocation of a specific premessenger ribonucleoprotein particle through the nuclear pore studied with electron microscope tomography. Cell. 1992 May 15;69(4):605–613. doi: 10.1016/0092-8674(92)90224-z. [DOI] [PubMed] [Google Scholar]
  27. Mehlin H., Skoglund U., Daneholt B. Transport of Balbiani ring granules through nuclear pores in Chironomus tentans. Exp Cell Res. 1991 Mar;193(1):72–77. doi: 10.1016/0014-4827(91)90539-7. [DOI] [PubMed] [Google Scholar]
  28. Neuman de Vegvar H. E., Dahlberg J. E. Nucleocytoplasmic transport and processing of small nuclear RNA precursors. Mol Cell Biol. 1990 Jul;10(7):3365–3375. doi: 10.1128/mcb.10.7.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Newmeyer D. D., Forbes D. J. Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation. Cell. 1988 Mar 11;52(5):641–653. doi: 10.1016/0092-8674(88)90402-3. [DOI] [PubMed] [Google Scholar]
  30. Newmeyer D. D. The nuclear pore complex and nucleocytoplasmic transport. Curr Opin Cell Biol. 1993 Jun;5(3):395–407. doi: 10.1016/0955-0674(93)90003-9. [DOI] [PubMed] [Google Scholar]
  31. Ofverstedt L. G., Zhang K., Tapio S., Skoglund U., Isaksson L. A. Starvation in vivo for aminoacyl-tRNA increases the spatial separation between the two ribosomal subunits. Cell. 1994 Nov 18;79(4):629–638. doi: 10.1016/0092-8674(94)90548-7. [DOI] [PubMed] [Google Scholar]
  32. Ohno M., Kataoka N., Shimura Y. A nuclear cap binding protein from HeLa cells. Nucleic Acids Res. 1990 Dec 11;18(23):6989–6995. doi: 10.1093/nar/18.23.6989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Olins A. L., Olins D. E., Franke W. W. Stereo-electron microscopy of nucleoli, Balbiani rings and endoplasmic reticulum in Chironomus salivary gland cells. Eur J Cell Biol. 1980 Oct;22(2):714–723. [PubMed] [Google Scholar]
  34. Paine P. L., Moore L. C., Horowitz S. B. Nuclear envelope permeability. Nature. 1975 Mar 13;254(5496):109–114. doi: 10.1038/254109a0. [DOI] [PubMed] [Google Scholar]
  35. Panté N., Aebi U. The nuclear pore complex. J Cell Biol. 1993 Sep;122(5):977–984. doi: 10.1083/jcb.122.5.977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Patzelt E., Blaas D., Kuechler E. CAP binding proteins associated with the nucleus. Nucleic Acids Res. 1983 Sep 10;11(17):5821–5835. doi: 10.1093/nar/11.17.5821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Radermacher M. Three-dimensional reconstruction of single particles from random and nonrandom tilt series. J Electron Microsc Tech. 1988 Aug;9(4):359–394. doi: 10.1002/jemt.1060090405. [DOI] [PubMed] [Google Scholar]
  38. Richardson W. D., Mills A. D., Dilworth S. M., Laskey R. A., Dingwall C. Nuclear protein migration involves two steps: rapid binding at the nuclear envelope followed by slower translocation through nuclear pores. Cell. 1988 Mar 11;52(5):655–664. doi: 10.1016/0092-8674(88)90403-5. [DOI] [PubMed] [Google Scholar]
  39. Rozen F., Sonenberg N. Identification of nuclear cap specific proteins in HeLa cells. Nucleic Acids Res. 1987 Aug 25;15(16):6489–6500. doi: 10.1093/nar/15.16.6489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scheer U., Dabauvalle M. C., Merkert H., Benevente R. The nuclear envelope and the organization of the pore complexes. Cell Biol Int Rep. 1988 Sep;12(9):669–689. doi: 10.1016/0309-1651(88)90083-5. [DOI] [PubMed] [Google Scholar]
  41. Schmekel K., Wahrman J., Skoglund U., Daneholt B. The central region of the synaptonemal complex in Blaps cribrosa studied by electron microscope tomography. Chromosoma. 1993 Dec;102(10):669–681. doi: 10.1007/BF00650893. [DOI] [PubMed] [Google Scholar]
  42. Schröder H. C., Bachmann M., Diehl-Seifert B., Müller W. E. Transport of mRNA from nucleus to cytoplasm. Prog Nucleic Acid Res Mol Biol. 1987;34:89–142. doi: 10.1016/s0079-6603(08)60494-8. [DOI] [PubMed] [Google Scholar]
  43. Skoglund U., Andersson K., Björkroth B., Lamb M. M., Daneholt B. Visualization of the formation and transport of a specific hnRNP particle. Cell. 1983 Oct;34(3):847–855. doi: 10.1016/0092-8674(83)90542-1. [DOI] [PubMed] [Google Scholar]
  44. Skoglund U., Andersson K., Strandberg B., Daneholt B. Three-dimensional structure of a specific pre-messenger RNP particle established by electron microscope tomography. Nature. 1986 Feb 13;319(6054):560–564. doi: 10.1038/319560a0. [DOI] [PubMed] [Google Scholar]
  45. Stevens B. J., Swift H. RNA transport from nucleus to cytoplasm in Chironomus salivary glands. J Cell Biol. 1966 Oct;31(1):55–77. doi: 10.1083/jcb.31.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stewart M. Nuclear pore structure and function. Semin Cell Biol. 1992 Aug;3(4):267–277. doi: 10.1016/1043-4682(92)90028-t. [DOI] [PubMed] [Google Scholar]
  47. Sukegawa J., Blobel G. A nuclear pore complex protein that contains zinc finger motifs, binds DNA, and faces the nucleoplasm. Cell. 1993 Jan 15;72(1):29–38. doi: 10.1016/0092-8674(93)90047-t. [DOI] [PubMed] [Google Scholar]
  48. Tobian J. A., Drinkard L., Zasloff M. tRNA nuclear transport: defining the critical regions of human tRNAimet by point mutagenesis. Cell. 1985 Dec;43(2 Pt 1):415–422. doi: 10.1016/0092-8674(85)90171-0. [DOI] [PubMed] [Google Scholar]
  49. Wickens M. P., Gurdon J. B. Post-transcriptional processing of simian virus 40 late transcripts in injected frog oocytes. J Mol Biol. 1983 Jan 5;163(1):1–26. doi: 10.1016/0022-2836(83)90027-x. [DOI] [PubMed] [Google Scholar]
  50. Wieslander L. The Balbiani ring multigene family: coding repetitive sequences and evolution of a tissue-specific cell function. Prog Nucleic Acid Res Mol Biol. 1994;48:275–313. doi: 10.1016/s0079-6603(08)60858-2. [DOI] [PubMed] [Google Scholar]
  51. Wilken N., Kossner U., Senécal J. L., Scheer U., Dabauvalle M. C. Nup180, a novel nuclear pore complex protein localizing to the cytoplasmic ring and associated fibrils. J Cell Biol. 1993 Dec;123(6 Pt 1):1345–1354. doi: 10.1083/jcb.123.6.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zasloff M. tRNA transport from the nucleus in a eukaryotic cell: carrier-mediated translocation process. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6436–6440. doi: 10.1073/pnas.80.21.6436. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES