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. 1981 Nov;78(11):6944–6948. doi: 10.1073/pnas.78.11.6944

Differential phosphorylation of nuclear nonhistone high mobility group proteins HMG 14 and HMG 17 during the cell cycle.

J S Bhorjee
PMCID: PMC349169  PMID: 6458819

Abstract

The phosphorylation of the high-mobility group (HMG) proteins at different stages of the cell cycle was studied in synchronized HeLa cells. HMG proteins were extracted and analyzed by NaDodSO4/polyacrylamide gel electrophoresis. Although the molecular weight distribution of HMGs remains unchanged, their total amounts increase by as much as 20-25% in the G1 and S phases when compared with amounts in G2. However, the most significant finding is that there is a 7-fold increase of 32P incorporation into HMG 14 in the G2 phase compared with that in G1, and a 2-fold increase of 32P incorporation into HMG 17 in early S phase relative to the incorporation in the G1 and G2 stages. In contrast, HMG 1 and HMG 2 are not phosphorylated. The clear demonstration of differential phosphorylation of HMG 14 and 17 at specific stages of the cell cycle warrants a serious consideration of their role in tissue-specific maintenance of the altered chromatin structure characteristic of potentially active or actively transcribed chromatin domains.

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Selected References

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

  1. Bhorjee J. S., Pederson T. Chromatin: its isolation from cultured mammalian cells with particular reference to contamination by nuclear ribnucleoprotein particles. Biochemistry. 1973 Jul 3;12(14):2766–2773. doi: 10.1021/bi00738a033. [DOI] [PubMed] [Google Scholar]
  2. Bhorjee J. S., Pederson T. Nonhistone chromosomal proteins in synchronized HeLa cells. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3345–3349. doi: 10.1073/pnas.69.11.3345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bustin M., Hopkins R. B., Isenberg I. Immunological relatedness of high mobility group chromosomal proteins from calf thymus. J Biol Chem. 1978 Mar 10;253(5):1694–1699. [PubMed] [Google Scholar]
  4. Caplan A., Ord M. G., Stocken L. A. Chromatin structure through the cell cycle. Studies with regeneration rat liver. Biochem J. 1978 Aug 15;174(2):475–483. doi: 10.1042/bj1740475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chambon P. Summary: the molecular biology of the eukaryotic genome is coming of age. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):1209–1234. doi: 10.1101/sqb.1978.042.01.122. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  8. Felsenfeld G. Chromatin. Nature. 1978 Jan 12;271(5641):115–122. doi: 10.1038/271115a0. [DOI] [PubMed] [Google Scholar]
  9. Gazit B., Panet A., Cedar H. Reconstitution of a deoxyribonuclease I-sensitive structure on active genes. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1787–1790. doi: 10.1073/pnas.77.4.1787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gershey E. L., Kleinsmith L. J. Phosphorylation of nuclear proteins in avian erythrocytes. Biochim Biophys Acta. 1969 Dec 23;194(2):519–525. doi: 10.1016/0005-2795(69)90113-5. [DOI] [PubMed] [Google Scholar]
  11. Goodwin G. H., Johns E. W. Are the high mobility group non-histone chromosomal proteins associated with 'active' chromatin? Biochim Biophys Acta. 1978 Jun 22;519(1):279–284. doi: 10.1016/0005-2787(78)90081-3. [DOI] [PubMed] [Google Scholar]
  12. Goodwin G. H., Mathew C. G., Wright C. A., Venkov C. D., Johns E. W. Analysis of the high mobility group proteins associated with salt-soluble nucleosomes. Nucleic Acids Res. 1979 Dec 11;7(7):1815–1835. doi: 10.1093/nar/7.7.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodwin G. H., Nicolas R. H., Johns E. W. An improved large scale fractionation of high mobility group non-histone chromatin proteins. Biochim Biophys Acta. 1975 Oct 20;405(2):280–291. doi: 10.1016/0005-2795(75)90094-x. [DOI] [PubMed] [Google Scholar]
  14. Goodwin G. H., Sanders C., Johns E. W. A new group of chromatin-associated proteins with a high content of acidic and basic amino acids. Eur J Biochem. 1973 Sep 21;38(1):14–19. doi: 10.1111/j.1432-1033.1973.tb03026.x. [DOI] [PubMed] [Google Scholar]
  15. Goodwin G. H., Walker J. M., Johns E. W. Studies on the degradation of high mobility group non-histone chromosomal proteins. Biochim Biophys Acta. 1978 Jun 22;519(1):233–242. doi: 10.1016/0005-2787(78)90076-x. [DOI] [PubMed] [Google Scholar]
  16. Gordon J. S., Rosenfeld B. I., Kaufman R., Williams D. L. Evidence for a quantitative tissue-specific distribution of high mobility group chromosomal proteins. Biochemistry. 1980 Sep 16;19(19):4395–4402. doi: 10.1021/bi00560a003. [DOI] [PubMed] [Google Scholar]
  17. Gurley L. R., Walters R. A., Tobey R. A. Sequential phsophorylation of histone subfractions in the Chinese hamster cell cycle. J Biol Chem. 1975 May 25;250(10):3936–3944. [PubMed] [Google Scholar]
  18. Jackson J. B., Pollock J. M., Jr, Rill R. L. Chromatin fractionation procedure that yields nucleosomes containing near-stoichiometric amounts of high mobility group nonhistone chromosomal proteins. Biochemistry. 1979 Aug 21;18(17):3739–3748. doi: 10.1021/bi00584a015. [DOI] [PubMed] [Google Scholar]
  19. Karn J., Johnson E. M., Vidali G., Allfrey V. G. Differential phosphorylation and turnover of nuclear acidic proteins during the cell cycle of synchronized HeLa cells. J Biol Chem. 1974 Feb 10;249(3):667–677. [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. 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]
  22. Levy W B., Wong N. C., Dixon G. H. Selective association of the trout-specific H6 protein with chromatin regions susceptible to DNase I and DNase II: possible location of HMG-T in the spacer region between core nucleosomes. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2810–2814. doi: 10.1073/pnas.74.7.2810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mathew C. G., Goodwin G. H., Johns E. W. Studies on the association of the high mobility group non-histone chromatin proteins with isolated nucleosomes. Nucleic Acids Res. 1979 Jan;6(1):167–179. doi: 10.1093/nar/6.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mitchison J. M. Enzyme synthesis in synchronous cultures. Science. 1969 Aug 15;165(3894):657–663. doi: 10.1126/science.165.3894.657. [DOI] [PubMed] [Google Scholar]
  25. Pederson T., Bhorjee J. S. Evidence for a role of RNA in eukaryotic chromosome structure. Metabolically stable, small nuclear RNA species are covalently linked to chromosomal DNA in HeLa cells. J Mol Biol. 1979 Mar 15;128(4):451–480. doi: 10.1016/0022-2836(79)90288-2. [DOI] [PubMed] [Google Scholar]
  26. Puck T. T. Phasing, Mitotic Delay, and Chromosomal Aberrations in Mammalian Cells. Science. 1964 May 1;144(3618):565–566. doi: 10.1126/science.144.3618.565-c. [DOI] [PubMed] [Google Scholar]
  27. Saffer J. D., Glazer R. I. The phosphorylation of high mobility group proteins 14 and 17 from Ehrlich ascites and L1210 in vitro. Biochem Biophys Res Commun. 1980 Apr 29;93(4):1280–1285. doi: 10.1016/0006-291x(80)90628-2. [DOI] [PubMed] [Google Scholar]
  28. Sandeen G., Wood W. I., Felsenfeld G. The interaction of high mobility proteins HMG14 and 17 with nucleosomes. Nucleic Acids Res. 1980 Sep 11;8(17):3757–3778. doi: 10.1093/nar/8.17.3757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sotirov N., Johns E. W. Quantitative differences in the content of the histone f2c between chicken erythrocytes and erythroblasts. Exp Cell Res. 1972 Jul;73(1):13–16. doi: 10.1016/0014-4827(72)90095-x. [DOI] [PubMed] [Google Scholar]
  30. Stalder J., Larsen A., Engel J. D., Dolan M., Groudine M., Weintraub H. Tissue-specific DNA cleavages in the globin chromatin domain introduced by DNAase I. Cell. 1980 Jun;20(2):451–460. doi: 10.1016/0092-8674(80)90631-5. [DOI] [PubMed] [Google Scholar]
  31. Sterner R., Boffa L. C., Vidali G. Comparative structural analysis of high mobility group proteins from a variety of sources. Evidence for a high mobility group protein unique to avian erythrocyte nuclei. J Biol Chem. 1978 Jun 10;253(11):3830–3836. [PubMed] [Google Scholar]
  32. Vidali G., Boffa L. C., Allfrey V. G. Selective release of chromosomal proteins during limited DNAase 1 digestion of avian erythrocyte chromatin. Cell. 1977 Oct;12(2):409–415. doi: 10.1016/0092-8674(77)90117-9. [DOI] [PubMed] [Google Scholar]
  33. Watson D. C., Peters E. H., Dixon G. H. The purification, characterization and partial sequence determination of a trout testis non-histone protein, HMG-T. Eur J Biochem. 1977 Mar 15;74(1):53–60. doi: 10.1111/j.1432-1033.1977.tb11365.x. [DOI] [PubMed] [Google Scholar]
  34. Weisbrod S., Groudine M., Weintraub H. Interaction of HMG 14 and 17 with actively transcribed genes. Cell. 1980 Jan;19(1):289–301. doi: 10.1016/0092-8674(80)90410-9. [DOI] [PubMed] [Google Scholar]
  35. Weisbrod S., Weintraub H. Isolation of a subclass of nuclear proteins responsible for conferring a DNase I-sensitive structure on globin chromatin. Proc Natl Acad Sci U S A. 1979 Feb;76(2):630–634. doi: 10.1073/pnas.76.2.630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weisbrod S., Weintraub H. Isolation of actively transcribed nucleosomes using immobilized HMG 14 and 17 and an analysis of alpha-globin chromatin. Cell. 1981 Feb;23(2):391–400. doi: 10.1016/0092-8674(81)90134-3. [DOI] [PubMed] [Google Scholar]
  37. Wu C. The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature. 1980 Aug 28;286(5776):854–860. doi: 10.1038/286854a0. [DOI] [PubMed] [Google Scholar]

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