Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Aug 1;103(2):465–474. doi: 10.1083/jcb.103.2.465

Dynamic behavior of histone H1 microinjected into HeLa cells

PMCID: PMC2113811  PMID: 3733875

Abstract

Histone H1 was purified from bovine thymus and radiolabeled with tritium by reductive methylation or with 125I using chloramine-T. Red blood cell-mediated microinjection was then used to introduce the labeled H1 molecules into HeLa cells synchronized in S phase. The injected H1 molecules rapidly entered HeLa nuclei, and a number of tests indicate that their association with chromatin was equivalent to that endogenous histone H1. The injected molecules copurified with HeLa cell nucleosomes, exhibited a half-life of approximately 100 h, and were hyperphosphorylated at mitosis. When injected HeLa cells were fused with mouse 3T3 fibroblasts less than 10% of the labeled H1 molecules migrated to mouse nuclei during the next 48 h. Thus, the intracellular behavior of histone H1 differs markedly from that of high mobility group proteins 1 and 2 (HMG1 and HMG2), which rapidly equilibrate between human and mouse nuclei after heterokaryon formation (Rechsteiner, M., and L. Kuehl, 1979, Cell, 16:901-908; Wu, L., M. Rechsteiner, and L. Kuehl, 1981, J. Cell Biol, 91: 488-496). Despite their slow rate of migration between nuclei, the injected H1 molecules were evenly distributed on mouse and human genomes soon after mitosis of HeLa-3T3 heterokaryons. These results suggest that although most histone H1 molecules are stably associated with interphase chromatin, they undergo extensive redistribution after mitosis.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Ajiro K., Borun T. W., Cohen L. H. Phosphorylation states of different histone 1 subtypes and their relationship to chromatin functions during the HeLa S-3 cell cycle. Biochemistry. 1981 Mar 17;20(6):1445–1454. doi: 10.1021/bi00509a007. [DOI] [PubMed] [Google Scholar]
  2. Brown D. D. The role of stable complexes that repress and activate eucaryotic genes. Cell. 1984 Jun;37(2):359–365. doi: 10.1016/0092-8674(84)90366-0. [DOI] [PubMed] [Google Scholar]
  3. Caron F., Thomas J. O. Exchange of histone H1 between segments of chromatin. J Mol Biol. 1981 Mar 15;146(4):513–537. doi: 10.1016/0022-2836(81)90045-0. [DOI] [PubMed] [Google Scholar]
  4. Delabar J. M. Nonrandom location of H1-H1 degree histones on chromatin of mouse liver and brain. J Biol Chem. 1985 Oct 15;260(23):12622–12628. [PubMed] [Google Scholar]
  5. Djondjurov L. P., Yancheva N. Y., Ivanova E. C. Histones of terminally differentiated cells undergo continuous turnover. Biochemistry. 1983 Aug 16;22(17):4095–4102. doi: 10.1021/bi00286a016. [DOI] [PubMed] [Google Scholar]
  6. Felsenfeld G., McGhee J. D. Structure of the 30 nm chromatin fiber. Cell. 1986 Feb 14;44(3):375–377. doi: 10.1016/0092-8674(86)90456-3. [DOI] [PubMed] [Google Scholar]
  7. Finch J. T., Klug A. Solenoidal model for superstructure in chromatin. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1897–1901. doi: 10.1073/pnas.73.6.1897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fischer S. G., Laemmli U. K. Cell cycle changes in Physarum polycephalum histone H1 phosphate: relationship to deoxyribonucleic acid binding and chromosome condensation. Biochemistry. 1980 May 13;19(10):2240–2246. doi: 10.1021/bi00551a038. [DOI] [PubMed] [Google Scholar]
  9. Glacy S. D. Pattern and time course of rhodamine-actin incorporation in cardiac myocytes. J Cell Biol. 1983 Apr;96(4):1164–1167. doi: 10.1083/jcb.96.4.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Green G. R., Poccia D. L. Phosphorylation of sea urchin sperm H1 and H2B histones precedes chromatin decondensation and H1 exchange during pronuclear formation. Dev Biol. 1985 Mar;108(1):235–245. doi: 10.1016/0012-1606(85)90026-0. [DOI] [PubMed] [Google Scholar]
  11. Gurley L. R., Walters R. A., Tobey R. A. The metabolism of histone fractions. IV. Synthesis of histones during the G1-phase of the mammalian life cycle. Arch Biochem Biophys. 1972 Feb;148(2):633–641. doi: 10.1016/0003-9861(72)90182-8. [DOI] [PubMed] [Google Scholar]
  12. Hannon R., Bateman E., Allan J., Harborne N., Gould H. Control of RNA polymerase binding to chromatin by variations in linker histone composition. J Mol Biol. 1984 Nov 25;180(1):131–149. doi: 10.1016/0022-2836(84)90434-0. [DOI] [PubMed] [Google Scholar]
  13. Hewish D. R., Burgoyne L. A. Chromatin sub-structure. The digestion of chromatin DNA at regularly spaced sites by a nuclear deoxyribonuclease. Biochem Biophys Res Commun. 1973 May 15;52(2):504–510. doi: 10.1016/0006-291x(73)90740-7. [DOI] [PubMed] [Google Scholar]
  14. Huang H. C., Cole R. D. The distribution of H1 histone is nonuniform in chromatin and correlates with different degrees of condensation. J Biol Chem. 1984 Nov 25;259(22):14237–14242. [PubMed] [Google Scholar]
  15. Igo-Kemenes T., Hörz W., Zachau H. G. Chromatin. Annu Rev Biochem. 1982;51:89–121. doi: 10.1146/annurev.bi.51.070182.000513. [DOI] [PubMed] [Google Scholar]
  16. Jackson V., Chalkley R. Histone synthesis and deposition in the G1 and S phases of hepatoma tissue culture cells. Biochemistry. 1985 Nov 19;24(24):6921–6930. doi: 10.1021/bi00345a026. [DOI] [PubMed] [Google Scholar]
  17. Jin Y. J., Cole R. D. H1 histone exchange is limited to particular regions of chromatin that differ in aggregation properties. J Biol Chem. 1986 Mar 5;261(7):3420–3427. [PubMed] [Google Scholar]
  18. Keith C. H., Feramisco J. R., Shelanski M. Direct visualization of fluorescein-labeled microtubules in vitro and in microinjected fibroblasts. J Cell Biol. 1981 Jan;88(1):234–240. doi: 10.1083/jcb.88.1.234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kornberg R. D. Chromatin structure: a repeating unit of histones and DNA. Science. 1974 May 24;184(4139):868–871. doi: 10.1126/science.184.4139.868. [DOI] [PubMed] [Google Scholar]
  20. Louters L., Chalkley R. Exchange of histones H1, H2A, and H2B in vivo. Biochemistry. 1985 Jun 18;24(13):3080–3085. doi: 10.1021/bi00334a002. [DOI] [PubMed] [Google Scholar]
  21. Louters L., Chalkley R. In vitro exchange of nucleosomal histones H2a and H2b. Biochemistry. 1984 Jan 31;23(3):547–552. doi: 10.1021/bi00298a023. [DOI] [PubMed] [Google Scholar]
  22. Manser T., Thacher T., Rechsteiner M. Arginine-rich histones do not exchange between human and mouse chromosomes in hybrid cells. Cell. 1980 Apr;19(4):993–1003. doi: 10.1016/0092-8674(80)90090-2. [DOI] [PubMed] [Google Scholar]
  23. McGhee J. D., Nickol J. M., Felsenfeld G., Rau D. C. Higher order structure of chromatin: orientation of nucleosomes within the 30 nm chromatin solenoid is independent of species and spacer length. Cell. 1983 Jul;33(3):831–841. doi: 10.1016/0092-8674(83)90025-9. [DOI] [PubMed] [Google Scholar]
  24. Olins A. L., Olins D. E. Spheroid chromatin units (v bodies). Science. 1974 Jan 25;183(4122):330–332. doi: 10.1126/science.183.4122.330. [DOI] [PubMed] [Google Scholar]
  25. Panyim S., Chalkley R. High resolution acrylamide gel electrophoresis of histones. Arch Biochem Biophys. 1969 Mar;130(1):337–346. doi: 10.1016/0003-9861(69)90042-3. [DOI] [PubMed] [Google Scholar]
  26. Rechsteiner M., Kuehl L. Microinjection of the nonhistone chromosomal protein HMG1 into bovine fibroblasts and HeLa cells. Cell. 1979 Apr;16(4):901–908. doi: 10.1016/0092-8674(79)90105-3. [DOI] [PubMed] [Google Scholar]
  27. Renz M., Nehls P., Hozier J. Involvement of histone H1 in the organization of the chromosome fiber. Proc Natl Acad Sci U S A. 1977 May;74(5):1879–1883. doi: 10.1073/pnas.74.5.1879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Richmond T. J., Finch J. T., Rushton B., Rhodes D., Klug A. Structure of the nucleosome core particle at 7 A resolution. Nature. 1984 Oct 11;311(5986):532–537. doi: 10.1038/311532a0. [DOI] [PubMed] [Google Scholar]
  29. Sakagami H., Mitsui Y., Murota S., Yamada M. Effect of growth stage on histone H1 metabolism in human diploid fibroblasts. J Cell Physiol. 1982 Feb;110(2):213–218. doi: 10.1002/jcp.1041100216. [DOI] [PubMed] [Google Scholar]
  30. Schlissel M. S., Brown D. D. The transcriptional regulation of Xenopus 5s RNA genes in chromatin: the roles of active stable transcription complexes and histone H1. Cell. 1984 Jul;37(3):903–913. doi: 10.1016/0092-8674(84)90425-2. [DOI] [PubMed] [Google Scholar]
  31. Strätling W. H., Klingholz R. Supranucleosomal structure of chromatin: digestion by calcium/magnesium endonuclease proceeds via a discrete size class of particles with elevated stability. Biochemistry. 1981 Mar 3;20(5):1386–1392. doi: 10.1021/bi00508a054. [DOI] [PubMed] [Google Scholar]
  32. Tack B. F., Dean J., Eilat D., Lorenz P. E., Schechter A. N. Tritium labeling of proteins to high specific radioactivity by reduction methylation. J Biol Chem. 1980 Sep 25;255(18):8842–8847. [PubMed] [Google Scholar]
  33. Thoma F., Koller T. Influence of histone H1 on chromatin structure. Cell. 1977 Sep;12(1):101–107. doi: 10.1016/0092-8674(77)90188-x. [DOI] [PubMed] [Google Scholar]
  34. Thomas J. O., Rees C. Exchange of histones H1 and H5 between chromatin fragments. A preference of H5 for higher-order structures. Eur J Biochem. 1983 Jul 15;134(1):109–115. doi: 10.1111/j.1432-1033.1983.tb07538.x. [DOI] [PubMed] [Google Scholar]
  35. Weintraub H. Assembly and propagation of repressed and depressed chromosomal states. Cell. 1985 Oct;42(3):705–711. doi: 10.1016/0092-8674(85)90267-3. [DOI] [PubMed] [Google Scholar]
  36. Weintraub H., Flint S. J., Leffak I. M., Groudine M., Grainger R. M. The generation and propagation of variegated chromosome structures. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):401–407. doi: 10.1101/sqb.1978.042.01.042. [DOI] [PubMed] [Google Scholar]
  37. Weintraub H. Histone-H1-dependent chromatin superstructures and the suppression of gene activity. Cell. 1984 Aug;38(1):17–27. doi: 10.1016/0092-8674(84)90522-1. [DOI] [PubMed] [Google Scholar]
  38. Widom J., Klug A. Structure of the 300A chromatin filament: X-ray diffraction from oriented samples. Cell. 1985 Nov;43(1):207–213. doi: 10.1016/0092-8674(85)90025-x. [DOI] [PubMed] [Google Scholar]
  39. Woodcock C. L., Frado L. L., Rattner J. B. The higher-order structure of chromatin: evidence for a helical ribbon arrangement. J Cell Biol. 1984 Jul;99(1 Pt 1):42–52. doi: 10.1083/jcb.99.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Worcel A., Strogatz S., Riley D. Structure of chromatin and the linking number of DNA. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1461–1465. doi: 10.1073/pnas.78.3.1461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wu L., Rechsteiner M., Kuehl L. Comparative studies on microinjected high-mobility-group chromosomal proteins, HMG1 and HMG2. J Cell Biol. 1981 Nov;91(2 Pt 1):488–496. doi: 10.1083/jcb.91.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zentgraf H., Müller U., Franke W. W. Supranucleosomal organization of sea urchin sperm chromatin in regularly arranged 40 to 50 nm large granular subunits. Eur J Cell Biol. 1980 Feb;20(3):254–264. [PubMed] [Google Scholar]
  43. van der Westhuyzen D. R., Böhm E. L., von Holt C. Fractionation of chicken erythrocyte whole histone into the six main components by gel exclusion chromatography. Biochim Biophys Acta. 1974 Aug 8;359(2):341–345. doi: 10.1016/0005-2795(74)90233-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES