Search tips
Search criteria 


Logo of jcellbiolHomeEditorsContactInstructions for Authors
J Cell Biol. 1988 July 1; 107(1): 57–68.
PMCID: PMC2115165

A trypsin-sensitive receptor on membrane vesicles is required for nuclear envelope formation in vitro


The reformation of functioning organelles at the end of mitosis presents a problem in vesicle targeting. Using extracts made from Xenopus laevis frog eggs, we have studied in vitro the vesicles that reform the nuclear envelope. In the in vitro assay, nuclear envelope growth is linear with time. Furthermore, the final surface area of the nuclear envelopes formed is directly dependent upon the amount of membrane vesicles added to the assay. Egg membrane vesicles could be fractionated into two populations, only one of which was competent for nuclear envelope assembly. We found that vesicles active in nuclear envelope assembly contained markers (BiP and alpha-glucosidase II) characteristic of the endoplasmic reticulum (ER), but that the majority of ER-derived vesicles do not contribute to nuclear envelope size. This functional distinction between nuclear vesicles and ER-derived vesicles implies that nuclear vesicles are unique and possess at least one factor required for envelope assembly that is lacking in other vesicles. Consistent with this, treatment of vesicles with trypsin destroyed their ability to form a nuclear envelope; electron microscopic studies indicate that the trypsin-sensitive proteins is required for vesicles to bind to chromatin. However, the protease- sensitive component(s) is resistant to treatments that disrupt protein- protein interactions, such as high salt, EDTA, or low ionic strength solutions. We propose that an integral membrane protein, or protein tightly associated with the membrane, is critical for nuclear vesicle targeting or function.

Full Text

The Full Text of this article is available as a PDF (2.8M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Benavente R, Krohne G, Franke WW. Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis. Cell. 1985 May;41(1):177–190. [PubMed]
  • Bole DG, Hendershot LM, Kearney JF. Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas. J Cell Biol. 1986 May;102(5):1558–1566. [PMC free article] [PubMed]
  • Blow JJ, Laskey RA. Initiation of DNA replication in nuclei and purified DNA by a cell-free extract of Xenopus eggs. Cell. 1986 Nov 21;47(4):577–587. [PubMed]
  • Blow JJ, Watson JV. Nuclei act as independent and integrated units of replication in a Xenopus cell-free DNA replication system. EMBO J. 1987 Jul;6(7):1997–2002. [PubMed]
  • Brands R, Snider MD, Hino Y, Park SS, Gelboin HV, Rothman JE. Retention of membrane proteins by the endoplasmic reticulum. J Cell Biol. 1985 Nov;101(5 Pt 1):1724–1732. [PMC free article] [PubMed]
  • Burke B, Gerace L. A cell free system to study reassembly of the nuclear envelope at the end of mitosis. Cell. 1986 Feb 28;44(4):639–652. [PubMed]
  • Burns DM, Touster O. Purification and characterization of glucosidase II, an endoplasmic reticulum hydrolase involved in glycoprotein biosynthesis. J Biol Chem. 1982 Sep 10;257(17):9990–10000. [PubMed]
  • Byers TJ, Armstrong PB. Membrane protein redistribution during Xenopus first cleavage. J Cell Biol. 1986 Jun;102(6):2176–2184. [PMC free article] [PubMed]
  • Colman A, Jones EA, Heasman J. Meiotic maturation in Xenopus oocytes: a link between the cessation of protein secretion and the polarized disappearance of Golgi apparati. J Cell Biol. 1985 Jul;101(1):313–318. [PMC free article] [PubMed]
  • Doms RW, Helenius A, White J. Membrane fusion activity of the influenza virus hemagglutinin. The low pH-induced conformational change. J Biol Chem. 1985 Mar 10;260(5):2973–2981. [PubMed]
  • Dunphy WG, Newport JW. Mitosis-inducing factors are present in a latent form during interphase in the Xenopus embryo. J Cell Biol. 1988 Jun;106(6):2047–2056. [PMC free article] [PubMed]
  • Forbes DJ, Kirschner MW, Newport JW. Spontaneous formation of nucleus-like structures around bacteriophage DNA microinjected into Xenopus eggs. Cell. 1983 Aug;34(1):13–23. [PubMed]
  • Franke WW. Structure, biochemistry, and functions of the nuclear envelope. Int Rev Cytol. 1974;Suppl 4:71–236. [PubMed]
  • Franke WW. Nuclear lamins and cytoplasmic intermediate filament proteins: a growing multigene family. Cell. 1987 Jan 16;48(1):3–4. [PubMed]
  • Franke WW, Scheer U, Krohne G, Jarasch ED. The nuclear envelope and the architecture of the nuclear periphery. J Cell Biol. 1981 Dec;91(3 Pt 2):39s–50s. [PMC free article] [PubMed]
  • Gerace L, Blobel G. The nuclear envelope lamina is reversibly depolymerized during mitosis. Cell. 1980 Jan;19(1):277–287. [PubMed]
  • Gerace L, Comeau C, Benson M. Organization and modulation of nuclear lamina structure. J Cell Sci Suppl. 1984;1:137–160. [PubMed]
  • Hino Y, Rothman JE. Glucosidase II, a glycoprotein of the endoplasmic reticulum membrane. Proteolytic cleavage into enzymatically active fragments. Biochemistry. 1985 Jan 29;24(3):800–805. [PubMed]
  • Kessel RG. The structure and function of annulate lamellae: porous cytoplasmic and intranuclear membranes. Int Rev Cytol. 1983;82:181–303. [PubMed]
  • Laskey RA, Mills AD, Morris NR. Assembly of SV40 chromatin in a cell-free system from Xenopus eggs. Cell. 1977 Feb;10(2):237–243. [PubMed]
  • Lohka MJ, Maller JL. Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts. J Cell Biol. 1985 Aug;101(2):518–523. [PMC free article] [PubMed]
  • Lohka MJ, Masui Y. Formation in vitro of sperm pronuclei and mitotic chromosomes induced by amphibian ooplasmic components. Science. 1983 May 13;220(4598):719–721. [PubMed]
  • Lohka MJ, Masui Y. Roles of cytosol and cytoplasmic particles in nuclear envelope assembly and sperm pronuclear formation in cell-free preparations from amphibian eggs. J Cell Biol. 1984 Apr;98(4):1222–1230. [PMC free article] [PubMed]
  • Longo FJ. Derivation of the membrane comprising the male pronuclear envelope in inseminated sea urchin eggs. Dev Biol. 1976 Apr;49(2):347–368. [PubMed]
  • Lucocq JM, Brada D, Roth J. Immunolocalization of the oligosaccharide trimming enzyme glucosidase II. J Cell Biol. 1986 Jun;102(6):2137–2146. [PMC free article] [PubMed]
  • Lucocq JM, Pryde JG, Berger EG, Warren G. A mitotic form of the Golgi apparatus in HeLa cells. J Cell Biol. 1987 Apr;104(4):865–874. [PMC free article] [PubMed]
  • Maul GG. The nuclear and the cytoplasmic pore complex: structure, dynamics, distribution, and evolution. Int Rev Cytol Suppl. 1977;(6):75–186. [PubMed]
  • Miake-Lye R, Kirschner MW. Induction of early mitotic events in a cell-free system. Cell. 1985 May;41(1):165–175. [PubMed]
  • Munro S, Pelham HR. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. [PubMed]
  • Newmeyer DD, Forbes DJ. Nuclear import can be separated into distinct steps in vitro: nuclear pore binding and translocation. Cell. 1988 Mar 11;52(5):641–653. [PubMed]
  • Newmeyer DD, Finlay DR, Forbes DJ. In vitro transport of a fluorescent nuclear protein and exclusion of non-nuclear proteins. J Cell Biol. 1986 Dec;103(6 Pt 1):2091–2102. [PMC free article] [PubMed]
  • Newmeyer DD, Lucocq JM, Bürglin TR, De Robertis EM. Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation. EMBO J. 1986 Mar;5(3):501–510. [PubMed]
  • Newport J. Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. Cell. 1987 Jan 30;48(2):205–217. [PubMed]
  • Newport JW, Forbes DJ. The nucleus: structure, function, and dynamics. Annu Rev Biochem. 1987;56:535–565. [PubMed]
  • Newport J, Kirschner M. A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982 Oct;30(3):675–686. [PubMed]
  • Newport J, Spann T. Disassembly of the nucleus in mitotic extracts: membrane vesicularization, lamin disassembly, and chromosome condensation are independent processes. Cell. 1987 Jan 30;48(2):219–230. [PubMed]
  • Newport J, Spann T, Kanki J, Forbes D. The role of mitotic factors in regulating the timing of the midblastula transition in Xenopus. Cold Spring Harb Symp Quant Biol. 1985;50:651–656. [PubMed]
  • Pathak RK, Luskey KL, Anderson RG. Biogenesis of the crystalloid endoplasmic reticulum in UT-1 cells: evidence that newly formed endoplasmic reticulum emerges from the nuclear envelope. J Cell Biol. 1986 Jun;102(6):2158–2168. [PMC free article] [PubMed]
  • Pollard HB, Rojas E, Burns AL. Synexin and chromaffin granule membrane fusion. A novel "hydrophobic bridge" hypothesis for the driving and directing of the fusion process. Ann N Y Acad Sci. 1987;493:524–541. [PubMed]
  • Puddington L, Lively MO, Lyles DS. Role of the nuclear envelope in synthesis, processing, and transport of membrane glycoproteins. J Biol Chem. 1985 May 10;260(9):5641–5647. [PubMed]
  • Reymond CD, Gomer RH, Mehdy MC, Firtel RA. Developmental regulation of a Dictyostelium gene encoding a protein homologous to mammalian ras protein. Cell. 1984 Nov;39(1):141–148. [PubMed]
  • Richardson JC, Maddy AH. The polypeptides of rat liver nuclear envelope. II. Comparison of rat liver nuclear membrane polypeptides with those of the rough endoplasmic reticulum. J Cell Sci. 1980 Jun;43:269–277. [PubMed]
  • Sheehan MA, Mills AD, Sleeman AM, Laskey RA, Blow JJ. Steps in the assembly of replication-competent nuclei in a cell-free system from Xenopus eggs. J Cell Biol. 1988 Jan;106(1):1–12. [PMC free article] [PubMed]
  • Stafstrom JP, Staehelin LA. Are annulate lamellae in the Drosophila embryo the result of overproduction of nuclear pore components? J Cell Biol. 1984 Feb;98(2):699–708. [PMC free article] [PubMed]
  • Stewart C, Burke B. Teratocarcinoma stem cells and early mouse embryos contain only a single major lamin polypeptide closely resembling lamin B. Cell. 1987 Nov 6;51(3):383–392. [PubMed]
  • Stick R, Hausen P. Changes in the nuclear lamina composition during early development of Xenopus laevis. Cell. 1985 May;41(1):191–200. [PubMed]
  • Wolin SL, Krohne G, Kirschner MW. A new lamin in Xenopus somatic tissues displays strong homology to human lamin A. EMBO J. 1987 Dec 1;6(12):3809–3818. [PubMed]
  • Woodland HR, Adamson ED. The synthesis and storage of histones during the oogenesis of Xenopus laevis. Dev Biol. 1977 May;57(1):118–135. [PubMed]

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