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. 1982 Jan 22;10(2):735–748. doi: 10.1093/nar/10.2.735

Distribution of the core histones H2A.H2B.H3 and H4 during cell replication.

E Fowler, R Farb, S El-Saidy
PMCID: PMC326181  PMID: 7063413

Abstract

The distribution of newly synthesized core histones H2A, H2B, H3 and H4 relative to the DNA strand synthesized in the same generation has been examined in replicating Chinese Hamster ovary cells. Cells are grown for one generation in [14C]-lysine and thymidine, and then for one generation in [3H]-lysine and 5-bromodeoxyuridine (BrUdRib) and a further generation in unlabeled lysine and thymidine. This protocol produces equal amounts of unifilarly substituted and unsubstituted DNA. Monomer nucleosomes isolated from chromatin containing these two types of DNA can be distinguished by crosslinking with formaldehyde and banding to equilibrium in CsCl density gradients. The results indicate that the core histones are equally distributed between the two types of DNA. These findings are discussed in terms of current models for chromatin replication; they do not support any long term association of newly replicated histones with either the leading or lagging side of the replication fork.

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Selected References

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

  1. Bloom K. S., Anderson J. N. Conformation of ovalbumin and globin genes in chromatin during differential gene expression. J Biol Chem. 1979 Oct 25;254(20):10532–10539. [PubMed] [Google Scholar]
  2. Fareed G. C., Garon G. F., Salzman N. P. Origin and direction of simian virus 40 deoxyribonucleic acid replication. J Virol. 1972 Sep;10(3):484–491. doi: 10.1128/jvi.10.3.484-491.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Freedlender E. F., Taichman L., Smithies O. Nonrandom distribution of chromosomal proteins during cell replication. Biochemistry. 1977 May 3;16(9):1802–1808. doi: 10.1021/bi00628a006. [DOI] [PubMed] [Google Scholar]
  4. Gottesfeld J. M., Butler P. J. Structure of transcriptionally-active chromatin subunits. Nucleic Acids Res. 1977 Sep;4(9):3155–3173. doi: 10.1093/nar/4.9.3155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Griffin B. E., Fried M., Cowie A. Polyoma DNA: a physical map. Proc Natl Acad Sci U S A. 1974 May;71(5):2077–2081. doi: 10.1073/pnas.71.5.2077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HAM R. G. An improved nutrient solution for diploid Chinese hamster and human cell lines. Exp Cell Res. 1963 Feb;29:515–526. doi: 10.1016/s0014-4827(63)80014-2. [DOI] [PubMed] [Google Scholar]
  7. Hancock R. Assembly of new nucleosomal histones and new DNA into chromatin. Proc Natl Acad Sci U S A. 1978 May;75(5):2130–2134. doi: 10.1073/pnas.75.5.2130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hand R. Eucaryotic DNA: organization of the genome for replication. Cell. 1978 Oct;15(2):317–325. doi: 10.1016/0092-8674(78)90001-6. [DOI] [PubMed] [Google Scholar]
  9. Harland R. M., Laskey R. A. Regulated replication of DNA microinjected into eggs of Xenopus laevis. Cell. 1980 Oct;21(3):761–771. doi: 10.1016/0092-8674(80)90439-0. [DOI] [PubMed] [Google Scholar]
  10. Jackson V., Chalkley R. A new method for the isolation of replicative chromatin: selective deposition of histone on both new and old DNA. Cell. 1981 Jan;23(1):121–134. doi: 10.1016/0092-8674(81)90277-4. [DOI] [PubMed] [Google Scholar]
  11. Jackson V., Chalkley R. A reevaluation of new histone deposition on replicating chromatin. J Biol Chem. 1981 May 25;256(10):5095–5103. [PubMed] [Google Scholar]
  12. Jackson V., Chalkley R. Separation of newly synthesized nucleohistone by equilibrium centrifugation in cesium chloride. Biochemistry. 1974 Sep 10;13(19):3952–3956. doi: 10.1021/bi00716a021. [DOI] [PubMed] [Google Scholar]
  13. Jackson V., Granner D. K., Chalkley R. Deposition of histones onto replicating chromosomes. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4440–4444. doi: 10.1073/pnas.72.11.4440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jackson V., Granner D., Chalkley R. Deposition of histone onto the replicating chromosome: newly synthesized histone is not found near the replication fork. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2266–2269. doi: 10.1073/pnas.73.7.2266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Klempnauer K. H., Fanning E., Otto B., Knippers R. Maturation of newly replicated chromatin of simian virus 40 and its host cell. J Mol Biol. 1980 Feb 5;136(4):359–374. doi: 10.1016/0022-2836(80)90395-2. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  18. Leffak I. M., Grainger R., Weintraub H. Conservative assembly and segregation of nucleosomal histones. Cell. 1977 Nov;12(3):837–845. doi: 10.1016/0092-8674(77)90282-3. [DOI] [PubMed] [Google Scholar]
  19. Levy A., Jakob K. M. Nascent DNA in nucleosome like structures from chromatin. Cell. 1978 Jun;14(2):259–267. doi: 10.1016/0092-8674(78)90112-5. [DOI] [PubMed] [Google Scholar]
  20. McGhee J. D., Felsenfeld G. Nucleosome structure. Annu Rev Biochem. 1980;49:1115–1156. doi: 10.1146/annurev.bi.49.070180.005343. [DOI] [PubMed] [Google Scholar]
  21. McKnight S. L., Miller O. L., Jr Electron microscopic analysis of chromatin replication in the cellular blastoderm Drosophila melanogaster embryo. Cell. 1977 Nov;12(3):795–804. doi: 10.1016/0092-8674(77)90278-1. [DOI] [PubMed] [Google Scholar]
  22. Nathans D., Danna K. J. Specific origin in SV40 DNA replication. Nat New Biol. 1972 Apr 19;236(68):200–202. doi: 10.1038/newbio236200a0. [DOI] [PubMed] [Google Scholar]
  23. Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
  24. Peacock A. C., Dingman C. W. Resolution of multiple ribonucleic acid species by polyacrylamide gel electrophoresis. Biochemistry. 1967 Jun;6(6):1818–1827. doi: 10.1021/bi00858a033. [DOI] [PubMed] [Google Scholar]
  25. Pfeiffer W., Zachau H. G. Accessibility of expressed and non-expressed genes to a restriction nuclease. Nucleic Acids Res. 1980 Oct 24;8(20):4621–4638. doi: 10.1093/nar/8.20.4621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Riley D., Weintraub H. Conservative segregation of parental histones during replication in the presence of cycloheximide. Proc Natl Acad Sci U S A. 1979 Jan;76(1):328–332. doi: 10.1073/pnas.76.1.328. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rill R. L., Shaw B. R., Van Holde K. E. Isolation and characterization of chromatin subunits. Methods Cell Biol. 1978;18:69–103. doi: 10.1016/s0091-679x(08)60134-x. [DOI] [PubMed] [Google Scholar]
  28. Robberson D. L., Clayton D. A., Morrow J. F. Cleavage of replicating forms of mitochondrial DNA by EcoRI endonuclease. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4447–4451. doi: 10.1073/pnas.71.11.4447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Russev G., Tsanev R. Nonrandom segregation of histones during chromatin replication. Eur J Biochem. 1979 Jan 2;93(1):123–128. doi: 10.1111/j.1432-1033.1979.tb12801.x. [DOI] [PubMed] [Google Scholar]
  30. Seale R. L., Simpson R. T. Effects of cycloheximide on chromatin biosynthesis. J Mol Biol. 1975 May 25;94(3):479–501. doi: 10.1016/0022-2836(75)90216-8. [DOI] [PubMed] [Google Scholar]
  31. Seale R. L. Site of histone assembly. Prog Nucleic Acid Res Mol Biol. 1981;26:123–134. doi: 10.1016/s0079-6603(08)60400-6. [DOI] [PubMed] [Google Scholar]
  32. Seale R. L. Studies on the mode of segregation of histone nu bodies during replication in HeLa cells. Cell. 1976 Nov;9(3):423–429. doi: 10.1016/0092-8674(76)90087-8. [DOI] [PubMed] [Google Scholar]
  33. Seale R. L. Studies on the mode of segregation of histone nu bodies during replication in HeLa cells. Cell. 1976 Nov;9(3):423–429. doi: 10.1016/0092-8674(76)90087-8. [DOI] [PubMed] [Google Scholar]
  34. Seale R. L. Temporal relationships of chromatin protein synthesis, DNA synthesis, and assembly of deoxyribonucleoprotein. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2270–2274. doi: 10.1073/pnas.73.7.2270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Seidman M. M., Levine A. J., Weintraub H. The asymmetric segregation of parental nucleosomes during chrosome replication. Cell. 1979 Oct;18(2):439–449. doi: 10.1016/0092-8674(79)90063-1. [DOI] [PubMed] [Google Scholar]
  36. Senshu T., Fukuda M., Ohashi M. Preferential association of newly synthesized H3 and H4 histones with newly replicated DNA. J Biochem. 1978 Oct;84(4):985–988. doi: 10.1093/oxfordjournals.jbchem.a132213. [DOI] [PubMed] [Google Scholar]
  37. Stalder J., Groudine M., Dodgson J. B., Engel J. D., Weintraub H. Hb switching in chickens. Cell. 1980 Apr;19(4):973–980. doi: 10.1016/0092-8674(80)90088-4. [DOI] [PubMed] [Google Scholar]
  38. Truett M. A., Gall J. G. The replication of ribosomal DNA in the macronucleus of Tetrahymena. Chromosoma. 1977 Dec 6;64(4):295–303. doi: 10.1007/BF00294937. [DOI] [PubMed] [Google Scholar]
  39. Tsanev R., Sendov B. Possible molecular mechanism for cell differentiation in multicellular organisms. J Theor Biol. 1971 Feb;30(2):337–393. doi: 10.1016/0022-5193(71)90059-2. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Weintraub H., Palter K., Van Lente F. Histones H2a, H2b, H3, and H4 form a tetrameric complex in solutions of high salt. Cell. 1975 Sep;6(1):85–110. doi: 10.1016/0092-8674(75)90077-x. [DOI] [PubMed] [Google Scholar]
  42. Worcel A., Han S., Wong M. L. Assembly of newly replicated chromatin. Cell. 1978 Nov;15(3):969–977. doi: 10.1016/0092-8674(78)90280-5. [DOI] [PubMed] [Google Scholar]

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