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. 1983 Jan 25;11(2):387–401. doi: 10.1093/nar/11.2.387

Affinity of HMG17 for a mononucleosome is not influenced by the presence of ubiquitin-H2A semihistone but strongly depends on DNA fragment size.

P S Swerdlow, A Varshavsky
PMCID: PMC325721  PMID: 6298725

Abstract

We have used a two-dimensional (deoxyribonucleoprotein leads to DNA) electrophoretic binding assay to study the interaction of the purified high mobility group protein HMG17 with isolated HeLa mononucleosomes as a function of their DNA fragment size and the presence of ubiquitin-H2A semihistone. No significant differences between affinities of HMG17 for ubiquitinated and non-ubiquitinated core mononucleosomes were observed. In striking contrast, the apparent affinity of HMG17 for a mononucleosome increases more than 100-fold upon an increase of the length of the mononucleosomal DNA fragment by as few as 3 to 5 bp over the core DNA length (integral of 146 bp). We suggest that the magnitude of this effect is sufficient to explain the preferential binding of HMG17 in vitro to mononucleosomes derived from actively transcribed genes.

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Selected References

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  1. Albanese I., Weintraub H. Electrophoretic separation of a class of nucleosomes enriched in HMG 14 and 17 and actively transcribed globin genes. Nucleic Acids Res. 1980 Jun 25;8(12):2787–2805. doi: 10.1093/nar/8.12.2787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albright S. C., Wiseman J. M., Lange R. A., Garrard W. T. Subunit structures of different electrophoretic forms of nucleosomes. J Biol Chem. 1980 Apr 25;255(8):3673–3684. [PubMed] [Google Scholar]
  3. Barsoum J., Levinger L., Varshavsky A. On the chromatin structure of the amplified, transcriptionally active gene for dihydrofolate reductase in mouse cells. J Biol Chem. 1982 May 10;257(9):5274–5282. [PubMed] [Google Scholar]
  4. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Busch H., Goldknopf I. L. Ubiquitin - protein conjugates. Mol Cell Biochem. 1981 Nov 13;40(3):173–187. doi: 10.1007/BF00224611. [DOI] [PubMed] [Google Scholar]
  6. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Isackson P. J., Reeck G. R. Nonhistone chromatin proteins HMG-14 and HMG-17 bind preferentially to single-stranded DNA. Nucleic Acids Res. 1981 Aug 11;9(15):3779–3791. doi: 10.1093/nar/9.15.3779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Levinger L., Barsoum J., Varshavsky A. Two-dimensional hybridization mapping of nucleosomes. comparison of DNA and protein patterns. J Mol Biol. 1981 Mar 5;146(3):287–304. doi: 10.1016/0022-2836(81)90389-2. [DOI] [PubMed] [Google Scholar]
  11. Levinger L., Varshavsky A. High-resolution fractionation of nucleosomes: minor particles, "whiskers," and separation of mononucleosomes containing and lacking A24 semihistone. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3244–3248. doi: 10.1073/pnas.77.6.3244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Levinger L., Varshavsky A. Selective arrangement of ubiquitinated and D1 protein-containing nucleosomes within the Drosophila genome. Cell. 1982 Feb;28(2):375–385. doi: 10.1016/0092-8674(82)90355-5. [DOI] [PubMed] [Google Scholar]
  13. Mardian J. K., Paton A. E., Bunick G. J., Olins D. E. Nucleosome cores have two specific binding sites for nonhistone chromosomal proteins HMG 14 and HMG 17. Science. 1980 Sep 26;209(4464):1534–1536. doi: 10.1126/science.7433974. [DOI] [PubMed] [Google Scholar]
  14. Matsui S., Sandberg A. A., Negoro S., Seon B. K., Goldstein G. Isopeptidase: a novel eukaryotic enzyme that cleaves isopeptide bonds. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1535–1539. doi: 10.1073/pnas.79.5.1535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Paulson J. R. Sulfhydryl reagents prevent dephosphorylation and proteolysis of histones in isolated HeLa metaphase chromosomes. Eur J Biochem. 1980 Oct;111(1):189–197. doi: 10.1111/j.1432-1033.1980.tb06092.x. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Todd R. D., Garrard W. T. Two-dimensional electrophoretic analysis of polynucleosomes. J Biol Chem. 1977 Jul 10;252(13):4729–4738. [PubMed] [Google Scholar]
  20. Weisbrod S. T. Properties of active nucleosomes as revealed by HMG 14 and 17 chromatography. Nucleic Acids Res. 1982 Mar 25;10(6):2017–2042. doi: 10.1093/nar/10.6.2017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Wu R. S., Kohn K. W., Bonner W. M. Metabolism of ubiquitinated histones. J Biol Chem. 1981 Jun 10;256(11):5916–5920. [PubMed] [Google Scholar]
  24. Zweidler A. Resolution of histones by polyacrylamide gel electrophoresis in presence of nonionic detergents. Methods Cell Biol. 1978;17:223–233. [PubMed] [Google Scholar]

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