Abstract
A series of extraction procedures were applied to avian nuclei which allowed us to define three types of association of v-myc- and c-myc-encoded proteins with nuclei: (i) a major fraction (60 to 90%) which is retained in DNA- and RNA-depleted nuclei after low- and high-salt extraction, (ii) a small fraction (1%) released during nuclease digestion of DNA in intact nuclei in the presence of low-salt buffer, and (iii) a fraction of myc protein (less than 10%) extractable with salt or detergents and found to have affinity for both single- and double-stranded DNA. Immunofluorescence analysis with anti-myc peptide sera on cells extracted sequentially with nucleases and salts confirmed the idea that myc proteins were associated with a complex residual nuclear structure (matrix-lamin fraction) which also contained avian nuclear lamin protein. Dispersal of myc proteins into the cytoplasm was found to occur during mitosis. Both c-myc and v-myc proteins were associated with the matrix-lamin, suggesting that the function of myc may relate to nuclear structural organization.
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- Abrams H. D., Rohrschneider L. R., Eisenman R. N. Nuclear location of the putative transforming protein of avian myelocytomatosis virus. Cell. 1982 Jun;29(2):427–439. doi: 10.1016/0092-8674(82)90159-3. [DOI] [PubMed] [Google Scholar]
- Aelen J. M., Opstelten R. J., Wanka F. Organization of DNA replication in Physarum polycephalum. Attachment of origins of replicons and replication forks to the nuclear matrix. Nucleic Acids Res. 1983 Feb 25;11(4):1181–1195. doi: 10.1093/nar/11.4.1181. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alitalo K., Ramsay G., Bishop J. M., Pfeifer S. O., Colby W. W., Levinson A. D. Identification of nuclear proteins encoded by viral and cellular myc oncogenes. Nature. 1983 Nov 17;306(5940):274–277. doi: 10.1038/306274a0. [DOI] [PubMed] [Google Scholar]
- Berezney R., Coffey D. S. Nuclear matrix. Isolation and characterization of a framework structure from rat liver nuclei. J Cell Biol. 1977 Jun;73(3):616–637. doi: 10.1083/jcb.73.3.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berezney R., Coffey D. S. Nuclear protein matrix: association with newly synthesized DNA. Science. 1975 Jul 25;189(4199):291–293. doi: 10.1126/science.1145202. [DOI] [PubMed] [Google Scholar]
- Bister K., Ramsay G. M., Hayman M. J. Deletions within the transformation-specific RNA sequences of acute leukemia virus MC29 give rise to partially transformation-defective mutants. J Virol. 1982 Mar;41(3):754–766. doi: 10.1128/jvi.41.3.754-766.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bodnar J. W., Jones C. J., Coombs D. H., Pearson G. D., Ward D. C. Proteins tightly bound to HeLa cell DNA at nuclear matrix attachment sites. Mol Cell Biol. 1983 Sep;3(9):1567–1579. doi: 10.1128/mcb.3.9.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buckler-White A. J., Humphrey G. W., Pigiet V. Association of polyoma T antigen and DNA with the nuclear matrix from lytically infected 3T6 cells. Cell. 1980 Nov;22(1 Pt 1):37–46. doi: 10.1016/0092-8674(80)90152-x. [DOI] [PubMed] [Google Scholar]
- Campisi J., Gray H. E., Pardee A. B., Dean M., Sonenshein G. E. Cell-cycle control of c-myc but not c-ras expression is lost following chemical transformation. Cell. 1984 Feb;36(2):241–247. doi: 10.1016/0092-8674(84)90217-4. [DOI] [PubMed] [Google Scholar]
- Capco D. G., Wan K. M., Penman S. The nuclear matrix: three-dimensional architecture and protein composition. Cell. 1982 Jul;29(3):847–858. doi: 10.1016/0092-8674(82)90446-9. [DOI] [PubMed] [Google Scholar]
- Ciejek E. M., Tsai M. J., O'Malley B. W. Actively transcribed genes are associated with the nuclear matrix. Nature. 1983 Dec 8;306(5943):607–609. doi: 10.1038/306607a0. [DOI] [PubMed] [Google Scholar]
- Clark R., Lane D. P., Tjian R. Use of monoclonal antibodies as probes of simian virus 40 T antigen ATPase activity. J Biol Chem. 1981 Nov 25;256(22):11854–11858. [PubMed] [Google Scholar]
- Cook P. R., Lang J., Hayday A., Lania L., Fried M., Chiswell D. J., Wyke J. A. Active viral genes in transformed cells lie close to the nuclear cage. EMBO J. 1982;1(4):447–452. doi: 10.1002/j.1460-2075.1982.tb01189.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donner P., Bunte T., Greiser-Wilke I., Moelling K. Decreased DNA-binding ability of purified transformation-specific proteins from deletion mutants of the acute avian leukemia virus MC29. Proc Natl Acad Sci U S A. 1983 May;80(10):2861–2865. doi: 10.1073/pnas.80.10.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donner P., Greiser-Wilke I., Moelling K. Nuclear localization and DNA binding of the transforming gene product of avian myelocytomatosis virus. Nature. 1982 Mar 18;296(5854):262–269. doi: 10.1038/296262a0. [DOI] [PubMed] [Google Scholar]
- Eisenman R. N., Linial M., Groudine M., Shaikh R., Brown S., Neiman P. E. Recombination in the avian oncoviruses as a model for the generation of defective transforming viruses. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1235–1247. doi: 10.1101/sqb.1980.044.01.134. [DOI] [PubMed] [Google Scholar]
- Enrietto P. J., Payne L. N., Hayman M. J. A recovered avian myelocytomatosis virus that induces lymphomas in chickens: pathogenic properties and their molecular basis. Cell. 1983 Dec;35(2 Pt 1):369–379. doi: 10.1016/0092-8674(83)90170-8. [DOI] [PubMed] [Google Scholar]
- Feldman L. T., Nevins J. R. Localization of the adenovirus E1Aa protein, a positive-acting transcriptional factor, in infected cells infected cells. Mol Cell Biol. 1983 May;3(5):829–838. doi: 10.1128/mcb.3.5.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fisher P. A., Berrios M., Blobel G. Isolation and characterization of a proteinaceous subnuclear fraction composed of nuclear matrix, peripheral lamina, and nuclear pore complexes from embryos of Drosophila melanogaster. J Cell Biol. 1982 Mar;92(3):674–686. doi: 10.1083/jcb.92.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerace L., Blobel G. The nuclear envelope lamina is reversibly depolymerized during mitosis. Cell. 1980 Jan;19(1):277–287. doi: 10.1016/0092-8674(80)90409-2. [DOI] [PubMed] [Google Scholar]
- Gerace L., Blum A., Blobel G. Immunocytochemical localization of the major polypeptides of the nuclear pore complex-lamina fraction. Interphase and mitotic distribution. J Cell Biol. 1978 Nov;79(2 Pt 1):546–566. doi: 10.1083/jcb.79.2.546. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hann S. R., Eisenman R. N. Proteins encoded by the human c-myc oncogene: differential expression in neoplastic cells. Mol Cell Biol. 1984 Nov;4(11):2486–2497. doi: 10.1128/mcb.4.11.2486. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayman M. J. Avian acute leukemia viruses. Curr Top Microbiol Immunol. 1983;103:109–125. doi: 10.1007/978-3-642-68943-7_5. [DOI] [PubMed] [Google Scholar]
- Jost E., Johnson R. T. Nuclear lamina assembly, synthesis and disaggregation during the cell cycle in synchronized HeLa cells. J Cell Sci. 1981 Feb;47:25–53. doi: 10.1242/jcs.47.1.25. [DOI] [PubMed] [Google Scholar]
- Kelly K., Cochran B. H., Stiles C. D., Leder P. Cell-specific regulation of the c-myc gene by lymphocyte mitogens and platelet-derived growth factor. Cell. 1983 Dec;35(3 Pt 2):603–610. doi: 10.1016/0092-8674(83)90092-2. [DOI] [PubMed] [Google Scholar]
- Lebkowski J. S., Laemmli U. K. Non-histone proteins and long-range organization of HeLa interphase DNA. J Mol Biol. 1982 Apr 5;156(2):325–344. doi: 10.1016/0022-2836(82)90332-1. [DOI] [PubMed] [Google Scholar]
- Mariman E. C., van Eekelen C. A., Reinders R. J., Berns A. J., van Venrooij W. J. Adenoviral heterogeneous nuclear RNA is associated with the host nuclear matrix during splicing. J Mol Biol. 1982 Jan 5;154(1):103–119. doi: 10.1016/0022-2836(82)90420-x. [DOI] [PubMed] [Google Scholar]
- Okazaki W., Witter R. L., Romero C., Nazerian K., Sharma J. M., Fadly A., Ewert D. Induction of lymphoid leukosis transplant able tumours and the establishment of lymphoblastoid cell lines. Avian Pathol. 1980 Jul;9(3):311–329. doi: 10.1080/03079458008418416. [DOI] [PubMed] [Google Scholar]
- Palmieri S., Kahn P., Graf T. Quail embryo fibroblasts transformed by four v-myc-containing virus isolates show enhanced proliferation but are non tumorigenic. EMBO J. 1983;2(12):2385–2389. doi: 10.1002/j.1460-2075.1983.tb01750.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quinlan M. P., Chen L. B., Knipe D. M. The intranuclear location of a herpes simplex virus DNA-binding protein is determined by the status of viral DNA replication. Cell. 1984 Apr;36(4):857–868. doi: 10.1016/0092-8674(84)90035-7. [DOI] [PubMed] [Google Scholar]
- Ramsay G., Graf T., Hayman M. J. Mutants of avian myelocytomatosis virus with smaller gag gene-related proteins have an altered transforming ability. Nature. 1980 Nov 13;288(5787):170–172. doi: 10.1038/288170a0. [DOI] [PubMed] [Google Scholar]
- Razin S. V., Chernokhvostov V. V., Roodyn A. V., Zbarsky I. B., Georgiev G. P. Proteins tightly bound to DNA in the regions of DNA attachment to the skeletal structures of interphase nuclei and metaphase chromosomes. Cell. 1981 Nov;27(1 Pt 2):65–73. doi: 10.1016/0092-8674(81)90361-5. [DOI] [PubMed] [Google Scholar]
- Robinson S. I., Nelkin B. D., Vogelstein B. The ovalbumin gene is associated with the nuclear matrix of chicken oviduct cells. Cell. 1982 Jan;28(1):99–106. doi: 10.1016/0092-8674(82)90379-8. [DOI] [PubMed] [Google Scholar]
- Royer-Pokora B., Beug H., Claviez M., Winkhardt H. J., Friis R. R., Graf T. Transformation parameters in chicken fibroblasts transformed by AEV and MC29 avian leukemia viruses. Cell. 1978 Apr;13(4):751–760. doi: 10.1016/0092-8674(78)90225-8. [DOI] [PubMed] [Google Scholar]
- Shaper J. H., Pardoll D. M., Kaufmann S. H., Barrack E. R., Vogelstein B., Coffey D. S. The relationship of the nuclear matrix to cellular structure and function. Adv Enzyme Regul. 1978;17:213–248. doi: 10.1016/0065-2571(79)90015-3. [DOI] [PubMed] [Google Scholar]
- Staufenbiel M., Deppert W. Different structural systems of the nucleus are targets for SV40 large T antigen. Cell. 1983 May;33(1):173–181. doi: 10.1016/0092-8674(83)90346-x. [DOI] [PubMed] [Google Scholar]
- Staufenbiel M., Deppert W. Preparation of nuclear matrices from cultured cells: subfractionation of nuclei in situ. J Cell Biol. 1984 May;98(5):1886–1894. doi: 10.1083/jcb.98.5.1886. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. S., Shepherd J. H., Mulvihill E. R., Palmiter R. D. Isolation of a nuclear ribonucleoprotein fraction from chick oviduct containing ovalbumin messenger RNA sequences. J Mol Biol. 1981 Aug 5;150(2):143–160. [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Verderame M. F., Kohtz D. S., Pollack R. E. 94,000- and 100,000-molecular-weight simian virus 40 T-antigens are associated with the nuclear matrix in transformed and revertant mouse cells. J Virol. 1983 May;46(2):575–583. doi: 10.1128/jvi.46.2.575-583.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weintraub H., Groudine M. Chromosomal subunits in active genes have an altered conformation. Science. 1976 Sep 3;193(4256):848–856. doi: 10.1126/science.948749. [DOI] [PubMed] [Google Scholar]