Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1977 Jan 1;72(1):194–208. doi: 10.1083/jcb.72.1.194

Nuclear matrix of HeLa S3 cells. Polypeptide composition during adenovirus infection and in phases of the cell cycle

PMCID: PMC2110990  PMID: 830654

Abstract

A subnuclear fraction has been isolated from HeLa S3 nuclei after treatment with high salt buffer, deoxyribonuclease, and dithiothreitol. This fraction retains the approximate size and shape of nuclei and resembles the nuclear matrix recently isolated from rat liver nuclei. Ultrastructural and biochemical analyses indicate that this structure consists of nonmembranous elements as well as some membranous elements. Its chemical composition is 87% protein, 12% phospholipid, 1% DNA, and 0.1% RNA by weight. The protein constituents are resolved in SDS- polyacrylamide slab gels into 30-35 distinguishable bands in the apparent molecular weight range of 14,000 - 200,000 with major peptides at 14,000 - 18,000 and 45,000 - 75,000. Analysis of newly synthesized polypeptides by cylindrical gel electrophoresis reveals another cluster in the 90,000-130,000 molecular weight range. Infection with adenovirus results in an altered polypeptide profile. Additional polypeptides with apparent molecular weights of 21,000, 23,000, and 92,000 become major components by 22 h after infection. Concomitantly, some peptides in the 45,000-75,000 mol wt range become less prominent. In synchronized cells the relative staining capacity of the six bands in the 45,000-75,000 mol wt range changes during the cell cycle. Synthesis of at least some matrix polypeptides occures in all phases of the cell cycle, although there is decreased synthesis in late S/G2. In the absence of protein synthesis after cell division, at least some polypeptides in the 45,000- 75,000 mol wt range survive nuclear dispersal and subsequent reformation during mitosis. The possible significance of this subnuclear structure with regard to structure-function relationships within the nucleus during virus replication and during the life cycle of the cell is discussed.

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

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

  1. Aaronson R. P., Blobel G. Isolation of nuclear pore complexes in association with a lamina. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1007–1011. doi: 10.1073/pnas.72.3.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Atkinson P. H., Summers D. F. Purification and properties of HeLa cell plasma membranes. J Biol Chem. 1971 Aug 25;246(16):5162–5175. [PubMed] [Google Scholar]
  3. BOOTSMA D., BUDKE L., VOS O. STUDIES ON SYNCHRONOUS DIVISION OF TISSUE CULTURE CELLS INITIATED BY EXCESS THYMIDINE. Exp Cell Res. 1964 Jan;33:301–309. doi: 10.1016/s0014-4827(64)81035-1. [DOI] [PubMed] [Google Scholar]
  4. Berezney R., Coffey D. S. Identification of a nuclear protein matrix. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1410–1417. doi: 10.1016/0006-291x(74)90355-6. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Chardonnet Y., Dales S. Early events in the interaction of adenoviruses with HeLa cells. 3. Relationship between an ATPase activity in nuclear envelopes and transfer of core material: a hypothesis. Virology. 1972 May;48(2):342–359. doi: 10.1016/0042-6822(72)90045-1. [DOI] [PubMed] [Google Scholar]
  7. Chardonnet Y., Dales S. Early events in the interaction of adenoviruses with HeLa cells. I. Penetration of type 5 and intracellular release of the DNA genome. Virology. 1970 Mar;40(3):462–477. doi: 10.1016/0042-6822(70)90189-3. [DOI] [PubMed] [Google Scholar]
  8. Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. I. Structural and chemical differentiation in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):73–96. doi: 10.1083/jcb.30.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Darlington R. W., Moss L. H., 3rd Herpesvirus envelopment. J Virol. 1968 Jan;2(1):48–55. doi: 10.1128/jvi.2.1.48-55.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  11. Erlandson R. A., de Harven E. The ultrastructure of synchronized HeLa cells. J Cell Sci. 1971 Mar;8(2):353–397. doi: 10.1242/jcs.8.2.353. [DOI] [PubMed] [Google Scholar]
  12. FOLCH J., ASCOLI I., LEES M., MEATH J. A., LeBARON N. Preparation of lipide extracts from brain tissue. J Biol Chem. 1951 Aug;191(2):833–841. [PubMed] [Google Scholar]
  13. Graham F. L., Abrahams P. J., Mulder C., Heijneker H. L., Warnaar S. O., De Vries F. A., Fiers W., Van Der Eb A. J. Studies on in vitro transformation by DNA and DNA fragments of human adenoviruses and simian virus 40. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):637–650. doi: 10.1101/sqb.1974.039.01.077. [DOI] [PubMed] [Google Scholar]
  14. Hodge L. D., Robbins E., Scharff M. D. Persistence of messenger RNA through mitosis in HeLa cells. J Cell Biol. 1969 Feb;40(2):497–507. doi: 10.1083/jcb.40.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hodge L. D., Scharff M. D. Effect of adenovirus on host cell DNA synthesis in synchronized cells. Virology. 1969 Apr;37(4):554–564. doi: 10.1016/0042-6822(69)90273-6. [DOI] [PubMed] [Google Scholar]
  16. Holtzman E., Smith I., Penman S. Electron microscopic studies of detergent-treated HeLa cell nuclei. J Mol Biol. 1966 May;17(1):131–135. doi: 10.1016/s0022-2836(66)80099-2. [DOI] [PubMed] [Google Scholar]
  17. Hubbard A. L., Cohn Z. A. The enzymatic iodination of the red cell membrane. J Cell Biol. 1972 Nov;55(2):390–405. doi: 10.1083/jcb.55.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kay R. R., Haines M. E., Johnston I. R. Late replication of the DNA associated with the nuclear membrane. FEBS Lett. 1971 Sep 1;16(4):233–236. doi: 10.1016/0014-5793(71)80358-7. [DOI] [PubMed] [Google Scholar]
  19. Lewis A. M., Jr, Rowe W. P. Studies on nondefective adenovirus-simian virus 40 hybrid viruses. I. A newly characterized simian virus 40 antigen induced by the Ad2+ND 1 virus. J Virol. 1971 Feb;7(2):189–197. doi: 10.1128/jvi.7.2.189-197.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maizel J. V., Jr, White D. O., Scharff M. D. The polypeptides of adenovirus. II. Soluble proteins, cores, top components and the structure of the virion. Virology. 1968 Sep;36(1):126–136. doi: 10.1016/0042-6822(68)90122-0. [DOI] [PubMed] [Google Scholar]
  21. OYAMA V. I., EAGLE H. Measurement of cell growth in tissue culture with a phenol reagent (folin-ciocalteau). Proc Soc Exp Biol Med. 1956 Feb;91(2):305–307. doi: 10.3181/00379727-91-22245. [DOI] [PubMed] [Google Scholar]
  22. Oho M., Riquetti P., Hudson B. G. Bovine renal glomerular basement membrane. Isolation and characterization of a glycoprotein component. J Biol Chem. 1975 Oct 10;250(19):7780–7787. [PubMed] [Google Scholar]
  23. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  24. ROBBINS E., MARCUS P. I. MITOTICALLY SYNCHRONIZED MAMMALIAN CELLS: A SIMPLE METHOD FOR OBTAINING LARGE POPULATIONS. Science. 1964 May 29;144(3622):1152–1153. doi: 10.1126/science.144.3622.1152. [DOI] [PubMed] [Google Scholar]
  25. Ray T. K., Skipski V. P., Barclay M., Essner E., Archibald F. M. Lipid composition of rat liver plasma membranes. J Biol Chem. 1969 Oct 25;244(20):5528–5536. [PubMed] [Google Scholar]
  26. Redman C. M. Proteolipid involvement in human erythrocyte membrane function. Biochim Biophys Acta. 1972 Sep 1;282(1):123–134. doi: 10.1016/0005-2736(72)90316-1. [DOI] [PubMed] [Google Scholar]
  27. Riley D. E., Keller J. M., Byers B. The isolation and characterization of nuclear ghosts from cultured HeLa cells. Biochemistry. 1975 Jul;14(13):3005–3013. doi: 10.1021/bi00684a033. [DOI] [PubMed] [Google Scholar]
  28. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  29. Simmons T., Heywood P., Hodge L. Nuclear envelope-associated resumption of RNA synthesis in late mitosis of HeLa cells. J Cell Biol. 1973 Oct;59(1):150–164. doi: 10.1083/jcb.59.1.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Skipski V. P., Peterson R. F., Barclay M. Quantitative analysis of phospholipids by thin-layer chromatography. Biochem J. 1964 Feb;90(2):374–378. doi: 10.1042/bj0900374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Williams C. A., Ockey C. H. Distribution of DNA replicator sites in mammalian nuclei after different methods of cell synchronization. Exp Cell Res. 1970 Dec;63(2):365–372. doi: 10.1016/0014-4827(70)90224-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES