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. 1991 Jan 2;112(2):225–235. doi: 10.1083/jcb.112.2.225

Vaults. III. Vault ribonucleoprotein particles open into flower-like structures with octagonal symmetry

PMCID: PMC2288824  PMID: 1988458

Abstract

The structure of rat liver vault ribonucleoprotein particles was examined using several different staining techniques in conjunction with EM and digestion with hydrolytic enzymes. Quantitative scanning transmission EM demonstrates that each vault particle has a total mass of 12.9 +/- 1 MD and contains two centers of mass, suggesting that each vault particle is a dimer. Freeze-etch reveals that each vault opens into delicate flower-like structures, in which eight rectangular petals are joined to a central ring, each by a thin hook. Vaults examined by negative stain and conventional transmission EM (CTEM) also reveal the flower-like structure. Trypsin treatment of vaults resulted exclusively in cleavage of the major vault protein (p104) and concurrently alters their structure as revealed by negative stain/CTEM, consistent with a localization of p104 to the flower petals. We propose a structural model that predicts the stoichiometry of vault proteins and RNA, defines vault dimer-monomer interactions, and describes two possible modes for unfolding of vaults into flowers. These highly dynamic structural variations are likely to play a role in vault function.

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Selected References

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  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. Interactions and structure of the nuclear pore complex revealed by cryo-electron microscopy. J Cell Biol. 1989 Sep;109(3):955–970. doi: 10.1083/jcb.109.3.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Akey C. W. Visualization of transport-related configurations of the nuclear pore transporter. Biophys J. 1990 Aug;58(2):341–355. doi: 10.1016/S0006-3495(90)82381-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Andrews D. W., Walter P., Ottensmeyer F. P. Evidence for an extended 7SL RNA structure in the signal recognition particle. EMBO J. 1987 Nov;6(11):3471–3477. doi: 10.1002/j.1460-2075.1987.tb02671.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boublik M., Robakis N., Hellmann W., Wall J. S. Electron microscopic study of free and ribosome bound rRNAs from E. coli. Eur J Cell Biol. 1982 Jun;27(2):177–184. [PubMed] [Google Scholar]
  6. Crowther R. A., Finch J. T., Pearse B. M. On the structure of coated vesicles. J Mol Biol. 1976 Jun 5;103(4):785–798. doi: 10.1016/0022-2836(76)90209-6. [DOI] [PubMed] [Google Scholar]
  7. Dreyfuss G. Structure and function of nuclear and cytoplasmic ribonucleoprotein particles. Annu Rev Cell Biol. 1986;2:459–498. doi: 10.1146/annurev.cb.02.110186.002331. [DOI] [PubMed] [Google Scholar]
  8. Franke W. W., Scheer U., Krohne G., Jarasch E. D. The nuclear envelope and the architecture of the nuclear periphery. J Cell Biol. 1981 Dec;91(3 Pt 2):39s–50s. doi: 10.1083/jcb.91.3.39s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gall J. G. Octagonal nuclear pores. J Cell Biol. 1967 Feb;32(2):391–399. doi: 10.1083/jcb.32.2.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerace L., Ottaviano Y., Kondor-Koch C. Identification of a major polypeptide of the nuclear pore complex. J Cell Biol. 1982 Dec;95(3):826–837. doi: 10.1083/jcb.95.3.826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goodenough U., Heuser J. Structural comparison of purified dynein proteins with in situ dynein arms. J Mol Biol. 1984 Dec 25;180(4):1083–1118. doi: 10.1016/0022-2836(84)90272-9. [DOI] [PubMed] [Google Scholar]
  12. Hanover J. A., Cohen C. K., Willingham M. C., Park M. K. O-linked N-acetylglucosamine is attached to proteins of the nuclear pore. Evidence for cytoplasmic and nucleoplasmic glycoproteins. J Biol Chem. 1987 Jul 15;262(20):9887–9894. [PubMed] [Google Scholar]
  13. Heuser J. E. Procedure for freeze-drying molecules adsorbed to mica flakes. J Mol Biol. 1983 Sep 5;169(1):155–195. doi: 10.1016/s0022-2836(83)80179-x. [DOI] [PubMed] [Google Scholar]
  14. Heuser J., Kirchhausen T. Deep-etch views of clathrin assemblies. J Ultrastruct Res. 1985 Jul-Aug;92(1-2):1–27. doi: 10.1016/0889-1605(85)90123-5. [DOI] [PubMed] [Google Scholar]
  15. Heuser J. Three-dimensional visualization of coated vesicle formation in fibroblasts. J Cell Biol. 1980 Mar;84(3):560–583. doi: 10.1083/jcb.84.3.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holt G. D., Snow C. M., Senior A., Haltiwanger R. S., Gerace L., Hart G. W. Nuclear pore complex glycoproteins contain cytoplasmically disposed O-linked N-acetylglucosamine. J Cell Biol. 1987 May;104(5):1157–1164. doi: 10.1083/jcb.104.5.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kedersha N. L., Miquel M. C., Bittner D., Rome L. H. Vaults. II. Ribonucleoprotein structures are highly conserved among higher and lower eukaryotes. J Cell Biol. 1990 Apr;110(4):895–901. doi: 10.1083/jcb.110.4.895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kedersha N. L., Rome L. H. Isolation and characterization of a novel ribonucleoprotein particle: large structures contain a single species of small RNA. J Cell Biol. 1986 Sep;103(3):699–709. doi: 10.1083/jcb.103.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kedersha N. L., Rome L. H. Preparative agarose gel electrophoresis for the purification of small organelles and particles. Anal Biochem. 1986 Jul;156(1):161–170. doi: 10.1016/0003-2697(86)90168-5. [DOI] [PubMed] [Google Scholar]
  20. Mandiyan V., Tumminia S., Wall J. S., Hainfeld J. F., Boublik M. Protein-induced conformational changes in 16 S ribosomal RNA during the initial assembly steps of the Escherichia coli 30 S ribosomal subunit. J Mol Biol. 1989 Nov 20;210(2):323–336. doi: 10.1016/0022-2836(89)90334-3. [DOI] [PubMed] [Google Scholar]
  21. Mosesson M. W., Hainfeld J., Wall J., Haschemeyer R. H. Identification and mass analysis of human fibrinogen molecules and their domains by scanning transmission electron microscopy. J Mol Biol. 1981 Dec 15;153(3):695–718. doi: 10.1016/0022-2836(81)90414-9. [DOI] [PubMed] [Google Scholar]
  22. Nierhaus K. H., Brimacombe R., Nowotny V., Pon C. L., Rheinberger H. J., Wittmann-Liebold B., Wittmann H. G. New aspects of structure, assembly, evolution, and function of ribosomes. Cold Spring Harb Symp Quant Biol. 1987;52:665–674. doi: 10.1101/sqb.1987.052.01.076. [DOI] [PubMed] [Google Scholar]
  23. Olson N. H., Baker T. S. Magnification calibration and the determination of spherical virus diameters using cryo-microscopy. Ultramicroscopy. 1989 Jul-Aug;30(3):281–297. doi: 10.1016/0304-3991(89)90057-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pardee J. D., Spudich J. A. Purification of muscle actin. Methods Enzymol. 1982;85(Pt B):164–181. doi: 10.1016/0076-6879(82)85020-9. [DOI] [PubMed] [Google Scholar]
  25. Prasad K., Alfsen A., Lippoldt R. E., Nandi P. K., Edelhoch H. Structural characterization of labeled clathrin and coated vesicles. Arch Biochem Biophys. 1984 Dec;235(2):403–410. doi: 10.1016/0003-9861(84)90213-3. [DOI] [PubMed] [Google Scholar]
  26. Reed R., Griffith J., Maniatis T. Purification and visualization of native spliceosomes. Cell. 1988 Jun 17;53(6):949–961. doi: 10.1016/s0092-8674(88)90489-8. [DOI] [PubMed] [Google Scholar]
  27. Reichelt R., Holzenburg A., Buhle E. L., Jr, Jarnik M., Engel A., Aebi U. Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components. J Cell Biol. 1990 Apr;110(4):883–894. doi: 10.1083/jcb.110.4.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Siegel V., Walter P. Each of the activities of signal recognition particle (SRP) is contained within a distinct domain: analysis of biochemical mutants of SRP. Cell. 1988 Jan 15;52(1):39–49. doi: 10.1016/0092-8674(88)90529-6. [DOI] [PubMed] [Google Scholar]
  29. Snow C. M., Senior A., Gerace L. Monoclonal antibodies identify a group of nuclear pore complex glycoproteins. J Cell Biol. 1987 May;104(5):1143–1156. doi: 10.1083/jcb.104.5.1143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Steven A. C., Hainfeld J. F., Wall J. S., Steer C. J. Mass distributions of coated vesicles isolated from liver and brain: analysis by scanning transmission electron microscopy. J Cell Biol. 1983 Dec;97(6):1714–1723. doi: 10.1083/jcb.97.6.1714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Unwin P. N., Milligan R. A. A large particle associated with the perimeter of the nuclear pore complex. J Cell Biol. 1982 Apr;93(1):63–75. doi: 10.1083/jcb.93.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vigers G. P., Crowther R. A., Pearse B. M. Three-dimensional structure of clathrin cages in ice. EMBO J. 1986 Mar;5(3):529–534. doi: 10.1002/j.1460-2075.1986.tb04242.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Walter P., Blobel G. Disassembly and reconstitution of signal recognition particle. Cell. 1983 Sep;34(2):525–533. doi: 10.1016/0092-8674(83)90385-9. [DOI] [PubMed] [Google Scholar]
  34. Wittmann H. G. Architecture of prokaryotic ribosomes. Annu Rev Biochem. 1983;52:35–65. doi: 10.1146/annurev.bi.52.070183.000343. [DOI] [PubMed] [Google Scholar]

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