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. 1974 Mar;71(3):644–647. doi: 10.1073/pnas.71.3.644

Chick-Erythrocyte Nucleus Reactivation in Heterokaryons: Suppression by Inhibitors of Proteolytic Enzymes

Zbigniew Darzynkiewicz *, Ewa Chelmicka-Szorc , Barry G W Arnason
PMCID: PMC388068  PMID: 4522779

Abstract

Reactivation of chick-erythrocyte nuclei in heterokaryons (obtained by Sendai virus-induced fusion of chick erythrocytes with HeLa cells) is suppressed by specific inhibitors of trypsin and trypsin-like enzymes. N-α-tosyl-L-lysyl-chloromethane and N-α-tosyl-L-arginine methylester inhibit erythrocyte nuclear enlargement and suppress RNA and DNA synthesis in nuclei of erythrocytes and HeLa cells in heterokaryons at concentrations that only minimally influence individual HeLa cells or HeLa homokaryons.

Although other unknown mechanisms of action cannot be formally excluded, the data are interpreted as fitting best with an intracellular site of action of the protease inhibitors studied, and as suggesting a role for cellular proteases in reactivation of chick-erythrocyte nuclei in heterokaryons.

Keywords: RNA and DNA synthesis, cell fusion, autoradiography, genome activation

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Selected References

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

  1. Bartley J., Chalkley R. Further studies of a thymus nucleohistone-associated protease. J Biol Chem. 1970 Sep 10;245(17):4286–4292. [PubMed] [Google Scholar]
  2. Berlowitz L., Pallotta D., Sibley C. H. Chromatin and histones: binding of tritiated actinomycin D to heterochromatin in mealy bugs. Science. 1969 Jun 27;164(3887):1527–1529. doi: 10.1126/science.164.3887.1527. [DOI] [PubMed] [Google Scholar]
  3. Bolund L., Darzynkiewicz Z., Ringertz N. R. Growth of hen erythrocyte nuclei undergoing reactivation in heterokaryons. Exp Cell Res. 1969 Aug;56(2):406–410. doi: 10.1016/0014-4827(69)90031-7. [DOI] [PubMed] [Google Scholar]
  4. Bolund L., Ringertz N. R., Harris H. Changes in the cytochemical properties of erythrocyte nuclei reactivated by cell fusion. J Cell Sci. 1969 Jan;4(1):71–87. doi: 10.1242/jcs.4.1.71. [DOI] [PubMed] [Google Scholar]
  5. Brachet J., Hulin N. Binding of tritiated actinomycin and cell differentiation. Nature. 1969 May 3;222(5192):481–482. doi: 10.1038/222481a0. [DOI] [PubMed] [Google Scholar]
  6. CAMERON I. L., PRESCOTT D. M. RNA and protein metabolism in the maturation of the nucleated chicken erythrocyte. Exp Cell Res. 1963 May;30:609–612. doi: 10.1016/0014-4827(63)90344-6. [DOI] [PubMed] [Google Scholar]
  7. Carlsson S. A., Moore G. P., Ringertz N. R. Nucleo-cytoplasmic protein migration during the activation of chick erythrocyte nuclei in heterokaryons. Exp Cell Res. 1973 Jan;76(1):234–241. doi: 10.1016/0014-4827(73)90441-2. [DOI] [PubMed] [Google Scholar]
  8. Darzynkiewicz Z., Bolund L., Ringertz N. R. Nucleoprotein changes and initiation of RNA synthesis in PHA stimulated lymphocytes. Exp Cell Res. 1969 Aug;56(2):418–424. doi: 10.1016/0014-4827(69)90033-0. [DOI] [PubMed] [Google Scholar]
  9. Darzynkiewicz Z., Chelmicka-Szorc E. Unscheduled DNA synthesis in hen erythrocyte nuclei reactivated in heterokaryons. Exp Cell Res. 1972 Sep;74(1):131–139. doi: 10.1016/0014-4827(72)90488-0. [DOI] [PubMed] [Google Scholar]
  10. Darzynkiewicz Z., Gledhill B. L. Uptake of tritium-labeled adenosine triphosphate by the cytoplasm of fixed interstitial cells from rat testes. J Histochem Cytochem. 1973 Dec;21(12):1035–1040. doi: 10.1177/21.12.1035. [DOI] [PubMed] [Google Scholar]
  11. Darzynkiewicz Z., Jacobson B. Effect of prednisolone on thymus lymphocytes. II. Studies on 3 H-actinomycin D Binding and RNA polymerase activity of isolate nuclei. Exp Cell Res. 1971 Jul;67(1):49–60. doi: 10.1016/0014-4827(71)90620-3. [DOI] [PubMed] [Google Scholar]
  12. Furlan M., Jericijo M., Suhar A. Purification and properties of a neutral protease from alf thymus nuclei. Biochim Biophys Acta. 1968 Aug 27;167(1):154–160. doi: 10.1016/0005-2744(68)90285-4. [DOI] [PubMed] [Google Scholar]
  13. Garrels J. I., Elgin S. C., Bonner J. A histone protease of rat liver chromatin. Biochem Biophys Res Commun. 1972 Jan 31;46(2):545–551. doi: 10.1016/s0006-291x(72)80173-6. [DOI] [PubMed] [Google Scholar]
  14. HARRIS H., WATKINS J. F. HYBRID CELLS DERIVED FROM MOUSE AND MAN: ARTIFICIAL HETEROKARYONS OF MAMMALIAN CELLS FROM DIFFERENT SPECIES. Nature. 1965 Feb 13;205:640–646. doi: 10.1038/205640a0. [DOI] [PubMed] [Google Scholar]
  15. Harris H. Behaviour of differentiated nuclei in heterokaryons of animal cells from different species. Nature. 1965 May 8;206(984):583–588. doi: 10.1038/206583a0. [DOI] [PubMed] [Google Scholar]
  16. Harris H., Sidebottom E., Grace D. M., Bramwell M. E. The expression of genetic information: a study with hybrid animal cells. J Cell Sci. 1969 Mar;4(2):499–525. doi: 10.1242/jcs.4.2.499. [DOI] [PubMed] [Google Scholar]
  17. Harris H. The reactivation of the red cell nucleus. J Cell Sci. 1967 Mar;2(1):23–32. doi: 10.1242/jcs.2.1.23. [DOI] [PubMed] [Google Scholar]
  18. Hirschhorn R., Grossman J., Troll W., Weissmann G. The effect of epsilon amino caproic acid and other inhibitors of proteolysis upon the response of human peripheral blood lymphocytes to phytohemagglutinin. J Clin Invest. 1971 Jun;50(6):1206–1217. doi: 10.1172/JCI106598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kernell A. M., Bolund L., Ringertz N. R. Chromatin changes during erythropoiesis. Exp Cell Res. 1971 Mar;65(1):1–6. doi: 10.1016/s0014-4827(71)80042-3. [DOI] [PubMed] [Google Scholar]
  20. Pederson T., Robbins E. Chromatin structure and the cell division cycle. Actinomycin binding in synchronized HeLa cells. J Cell Biol. 1972 Nov;55(2):322–327. doi: 10.1083/jcb.55.2.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ringertz N. R., Bolund L. Actinomycin binding capacity of deoxyribonucleoprotein. Biochim Biophys Acta. 1969 Jan 21;174(1):147–154. doi: 10.1016/0005-2787(69)90237-8. [DOI] [PubMed] [Google Scholar]
  22. Ringertz N. R., Carlsson S. A., Ege T., Bolund L. Detection of human and chick nuclear antigens in nuclei of chick erythrocytes during reactivation in heterokaryons with HeLa cells. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3228–3232. doi: 10.1073/pnas.68.12.3228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schneeberger E. E., Harris H. An ultrastructural study of inter-specific cell fusion induced by inactivated Sendai virus. J Cell Sci. 1966 Dec;1(4):401–406. doi: 10.1242/jcs.1.4.401. [DOI] [PubMed] [Google Scholar]

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