Abstract
The mechanism by which proteins accumulate in the cell nucleus is not yet known. Two alternative mechanisms are discussed here: (a) selective unidirectional entry of karyophilic proteins through the nuclear pores, and (b) free diffusion of all proteins through the nuclear pores and specific binding of nuclear proteins to nondiffusible components of the nucleoplasm. We present experiments designed to distinguish between these alternatives. After mechanical injury of the Xenopus oocyte nuclear envelope, nuclear proteins were detected in the cytoplasm by immunohistochemical methods. In a second approach, nuclei from X. borealis oocytes were isolated under oil, the nuclear envelopes were removed, and the pure nucleoplasm was injected into the vegetal pole of X. laevis oocytes. With immunohistochemical methods, it was found that each of five nuclear proteins rapidly diffuses out of the injected nucleoplasm into the surrounding cytoplasm. The subsequent transport and accumulation in the intact host nucleus could be shown for the nuclear protein N1 with the aid of a species-specific mAb that reacts only with X. borealis N1. Purified and iodinated nucleoplasmin was injected into the cytoplasm of Xenopus oocytes and its uptake into the nucleus was studied by biochemical methods.
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- Bonner W. M. Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins. J Cell Biol. 1975 Feb;64(2):421–430. doi: 10.1083/jcb.64.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark T. G., Merriam R. W. Diffusible and bound actin nuclei of Xenopus laevis oocytes. Cell. 1977 Dec;12(4):883–891. doi: 10.1016/0092-8674(77)90152-0. [DOI] [PubMed] [Google Scholar]
- Dabauvalle M. C., Franke W. W. Karyophilic proteins: polypeptides synthesized in vitro accumulate in the nucleus on microinjection into the cytoplasm of amphibian oocytes. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5302–5306. doi: 10.1073/pnas.79.17.5302. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davey J., Dimmock N. J., Colman A. Identification of the sequence responsible for the nuclear accumulation of the influenza virus nucleoprotein in Xenopus oocytes. Cell. 1985 Mar;40(3):667–675. doi: 10.1016/0092-8674(85)90215-6. [DOI] [PubMed] [Google Scholar]
- De Robertis E. M., Longthorne R. F., Gurdon J. B. Intracellular migration of nuclear proteins in Xenopus oocytes. Nature. 1978 Mar 16;272(5650):254–256. doi: 10.1038/272254a0. [DOI] [PubMed] [Google Scholar]
- De Robertis E. M. Nucleocytoplasmic segregation of proteins and RNAs. Cell. 1983 Apr;32(4):1021–1025. doi: 10.1016/0092-8674(83)90285-4. [DOI] [PubMed] [Google Scholar]
- Dingwall C., Laskey R. A. Protein import into the cell nucleus. Annu Rev Cell Biol. 1986;2:367–390. doi: 10.1146/annurev.cb.02.110186.002055. [DOI] [PubMed] [Google Scholar]
- Dingwall C., Sharnick S. V., Laskey R. A. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell. 1982 Sep;30(2):449–458. doi: 10.1016/0092-8674(82)90242-2. [DOI] [PubMed] [Google Scholar]
- Dreyer C., Hausen P. Two-dimensional gel analysis of the fate of oocyte nuclear proteins in the development of Xenopus laevis. Dev Biol. 1983 Dec;100(2):412–425. doi: 10.1016/0012-1606(83)90235-x. [DOI] [PubMed] [Google Scholar]
- Dreyer C., Wang Y. H., Hausen P. Immunological relationship between oocyte nuclear proteins of Xenopus laevis and X. borealis. Dev Biol. 1985 Mar;108(1):210–219. doi: 10.1016/0012-1606(85)90024-7. [DOI] [PubMed] [Google Scholar]
- Feldherr C. M., Ogburn J. A. Mechanism for the selection of nuclear polypeptides in Xenopus oocytes. II. Two-dimensional gel analysis. J Cell Biol. 1980 Dec;87(3 Pt 1):589–593. doi: 10.1083/jcb.87.3.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feldherr C. M., Pomerantz J. Mechanism for the selection of nuclear polypeptides in Xenopus oocytes. J Cell Biol. 1978 Jul;78(1):168–175. doi: 10.1083/jcb.78.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gurdon J. B. Injected nuclei in frog oocytes: fate, enlargement, and chromatin dispersal. J Embryol Exp Morphol. 1976 Dec;36(3):523–540. [PubMed] [Google Scholar]
- Gurdon J. B. Nuclear transplantation and the control of gene activity in animal development. Proc R Soc Lond B Biol Sci. 1970 Dec 1;176(1044):303–314. doi: 10.1098/rspb.1970.0050. [DOI] [PubMed] [Google Scholar]
- Hall M. N., Hereford L., Herskowitz I. Targeting of E. coli beta-galactosidase to the nucleus in yeast. Cell. 1984 Apr;36(4):1057–1065. doi: 10.1016/0092-8674(84)90055-2. [DOI] [PubMed] [Google Scholar]
- Kalderon D., Richardson W. D., Markham A. F., Smith A. E. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984 Sep 6;311(5981):33–38. doi: 10.1038/311033a0. [DOI] [PubMed] [Google Scholar]
- Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. A short amino acid sequence able to specify nuclear location. Cell. 1984 Dec;39(3 Pt 2):499–509. doi: 10.1016/0092-8674(84)90457-4. [DOI] [PubMed] [Google Scholar]
- Krohne G., Franke W. W. A major soluble acidic protein located in nuclei of diverse vertebrate species. Exp Cell Res. 1980 Sep;129(1):167–189. doi: 10.1016/0014-4827(80)90341-9. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Langone J. J. Applications of immobilized protein A in immunochemical techniques. J Immunol Methods. 1982 Dec 30;55(3):277–296. doi: 10.1016/0022-1759(82)90088-6. [DOI] [PubMed] [Google Scholar]
- Laskey R. A., Honda B. M., Mills A. D., Finch J. T. Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA. Nature. 1978 Oct 5;275(5679):416–420. doi: 10.1038/275416a0. [DOI] [PubMed] [Google Scholar]
- Mills A. D., Laskey R. A., Black P., De Robertis E. M. An acidic protein which assembles nucleosomes in vitro is the most abundant protein in Xenopus oocyte nuclei. J Mol Biol. 1980 May 25;139(3):561–568. doi: 10.1016/0022-2836(80)90148-5. [DOI] [PubMed] [Google Scholar]
- Neukirchen R. O., Schlosshauer B., Baars S., Jäckle H., Schwarz U. Two-dimensional protein analysis at high resolution on a microscale. J Biol Chem. 1982 Dec 25;257(24):15229–15234. [PubMed] [Google Scholar]
- Newmeyer D. D., Finlay D. R., Forbes D. J. 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. doi: 10.1083/jcb.103.6.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Newmeyer D. D., Lucocq J. M., Bürglin T. R., De Robertis E. M. Assembly in vitro of nuclei active in nuclear protein transport: ATP is required for nucleoplasmin accumulation. EMBO J. 1986 Mar;5(3):501–510. doi: 10.1002/j.1460-2075.1986.tb04239.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nilsson K., Mosbach K. Immobilization of ligands with organic sulfonyl chlorides. Methods Enzymol. 1984;104:56–69. doi: 10.1016/s0076-6879(84)04083-0. [DOI] [PubMed] [Google Scholar]
- Paine P. L., Austerberry C. F., Desjarlais L. J., Horowitz S. B. Protein loss during nuclear isolation. J Cell Biol. 1983 Oct;97(4):1240–1242. doi: 10.1083/jcb.97.4.1240. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Richardson W. D., Roberts B. L., Smith A. E. Nuclear location signals in polyoma virus large-T. Cell. 1986 Jan 17;44(1):77–85. doi: 10.1016/0092-8674(86)90486-1. [DOI] [PubMed] [Google Scholar]
- Sealy L., Cotten M., Chalkley R. Xenopus nucleoplasmin: egg vs. oocyte. Biochemistry. 1986 May 20;25(10):3064–3072. doi: 10.1021/bi00358a049. [DOI] [PubMed] [Google Scholar]
- Wallace R. A., Jared D. W., Dumont J. N., Sega M. W. Protein incorporation by isolated amphibian oocytes. 3. Optimum incubation conditions. J Exp Zool. 1973 Jun;184(3):321–333. doi: 10.1002/jez.1401840305. [DOI] [PubMed] [Google Scholar]
- Wedlich D., Dreyer C., Hausen P. Occurrence of a species-specific nuclear antigen in the germ line of Xenopus and its expression from paternal genes in hybrid frogs. Dev Biol. 1985 Mar;108(1):220–234. doi: 10.1016/0012-1606(85)90025-9. [DOI] [PubMed] [Google Scholar]
- Wickner W. T., Lodish H. F. Multiple mechanisms of protein insertion into and across membranes. Science. 1985 Oct 25;230(4724):400–407. doi: 10.1126/science.4048938. [DOI] [PubMed] [Google Scholar]