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. 1986 Dec 1;103(6):2083–2089. doi: 10.1083/jcb.103.6.2083

Monoclonal antibody specific for human nuclear proteins IEF 8Z30 and 8Z31 accumulates in the nucleus a few hours after cytoplasmic microinjection of cells expressing these proteins

PMCID: PMC2114576  PMID: 3782292

Abstract

A monoclonal antibody (mAB 1C4C10) that reacts specifically with human nuclear proteins IEF 8Z30 and 8Z31 (charge variants; HeLa protein catalogue number; Bravo, R., and J. E. Celis, 1982, Clin. Chem., 28:766- 781) has been microinjected into the cytoplasm of cultured cells that either express (primates) or lack these proteins (at least having similar molecular weights and pIs; other species), and its cellular localization has been determined by indirect immunofluorescence. Nuclear localization (nucleolar and nucleoplasmic) of the antibody was observed only in cells expressing these antigens, suggesting that a determinant present in IEF 8Z30 and 8Z31 is required for cytoplasm- nuclear translocation. Nuclear migration was not inhibited by cycloheximide, implying that these proteins may shuttle between nucleus and cytoplasm. The results assumed to support the signal rather than the free diffusion model are further supported by microinjection experiments using antibodies (proliferating cell nuclear antigen/cyclin, DNA) that react with nuclear components but do not recognize cytoplasmic antigens. Furthermore, they raise the possibility that some nonnuclear proteins may be transported to the nucleus by interacting with proteins harboring nuclear location signals.

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Selected References

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

  1. Bennett F. C., Busch H., Lischwe M. A., Yeoman L. C. Antibodies to a nucleolar protein are localized in the nucleolus after red blood cell-mediated microinjection. J Cell Biol. 1983 Nov;97(5 Pt 1):1566–1572. doi: 10.1083/jcb.97.5.1566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bravo R., Celis J. E. Up-dated catalogue of HeLa cell proteins: percentages and characteristics of the major cell polypeptides labeled with a mixture of 16 14C-labeled amino acids. Clin Chem. 1982 Apr;28(4 Pt 2):766–781. [PubMed] [Google Scholar]
  4. Bravo R., Fey S. J., Small J. V., Larsen P. M., Celis J. E. Coexistence of three major isoactins in a single sarcoma 180 cell. Cell. 1981 Jul;25(1):195–202. doi: 10.1016/0092-8674(81)90244-0. [DOI] [PubMed] [Google Scholar]
  5. Bravo R., Macdonald-Bravo H. Changes in the nuclear distribution of cyclin (PCNA) but not its synthesis depend on DNA replication. EMBO J. 1985 Mar;4(3):655–661. doi: 10.1002/j.1460-2075.1985.tb03679.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bravo R., Small J. V., Fey S. J., Larsen P. M., Celis J. E. Architecture and polypeptide composition of HeLa cytoskeletons. Modification of cytoarchitectural polypeptides during mitosis. J Mol Biol. 1982 Jan 5;154(1):121–143. doi: 10.1016/0022-2836(82)90421-1. [DOI] [PubMed] [Google Scholar]
  7. Celis J. E., Celis A. Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase. Proc Natl Acad Sci U S A. 1985 May;82(10):3262–3266. doi: 10.1073/pnas.82.10.3262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Celis J. E. Injection of tRNAs into somatic cells: search for in vivo systems to assay potential nonsense mutations in somatic cells. Brookhaven Symp Biol. 1977 May 12;(29):178–196. [PubMed] [Google Scholar]
  9. Celis J. E., Madsen P., Nielsen S., Celis A. Nuclear patterns of cyclin (PCNA) antigen distribution subdivide S-phase in cultured cells--some applications of PCNA antibodies. Leuk Res. 1986;10(3):237–249. doi: 10.1016/0145-2126(86)90021-4. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. 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]
  13. 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]
  14. 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]
  15. Lanford R. E., Butel J. S. Construction and characterization of an SV40 mutant defective in nuclear transport of T antigen. Cell. 1984 Jul;37(3):801–813. doi: 10.1016/0092-8674(84)90415-x. [DOI] [PubMed] [Google Scholar]
  16. Mathews M. B., Bernstein R. M., Franza B. R., Jr, Garrels J. I. Identity of the proliferating cell nuclear antigen and cyclin. Nature. 1984 May 24;309(5966):374–376. doi: 10.1038/309374a0. [DOI] [PubMed] [Google Scholar]
  17. Miyachi K., Fritzler M. J., Tan E. M. Autoantibody to a nuclear antigen in proliferating cells. J Immunol. 1978 Dec;121(6):2228–2234. [PubMed] [Google Scholar]
  18. Mose-Larsen P., Bravo R., Fey S. J., Small J. V., Celis J. E. Putative association of mitochondria with a subpopulation of intermediate-sized filaments in cultured human skin fibroblasts. Cell. 1982 Dec;31(3 Pt 2):681–692. doi: 10.1016/0092-8674(82)90323-3. [DOI] [PubMed] [Google Scholar]
  19. Paucha E., Kalderon D., Richardson W. D., Harvey R. W., Smith A. E. The abnormal location of cytoplasmic SV40 large T is not caused by failure to bind to DNA or to p53. EMBO J. 1985 Dec 1;4(12):3235–3240. doi: 10.1002/j.1460-2075.1985.tb04071.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Silver P. A., Keegan L. P., Ptashne M. Amino terminus of the yeast GAL4 gene product is sufficient for nuclear localization. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5951–5955. doi: 10.1073/pnas.81.19.5951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tsuneoka M., Imamoto N. S., Uchida T. Monoclonal antibody against non-histone chromosomal protein high mobility group 1 Co-migrates with high mobility group 1 into the nucleus. J Biol Chem. 1986 Feb 5;261(4):1829–1834. [PubMed] [Google Scholar]

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