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. 1993 Oct;12(10):3855–3864. doi: 10.1002/j.1460-2075.1993.tb06064.x

Deposition of chromosomal protein HMG-17 during replication affects the nucleosomal ladder and transcriptional potential of nascent chromatin.

M P Crippa 1, L Trieschmann 1, P J Alfonso 1, A P Wolffe 1, M Bustin 1
PMCID: PMC413669  PMID: 8404854

Abstract

A cell-free system from Xenopus eggs was used to study the role of chromosomal protein HMG-17 in the generation of the chromatin structure of transcriptionally active genes. Addition of HMG-17 protein to the extracts, which do not contain structural homologs of the HMG-14/-17 protein family, indicates the protein is incorporated into the nascent template during replication, prior to completion of chromatin assembly. The protein binds to and stabilizes the structure of the nucleosomal core thereby improving the apparent periodicity of the nucleosomal spacing of nascent chromatin. Assembly of HMG-17 into the nascent chromatin structure significantly increased the transcription potential of the 5S RNA gene and satellite I chromatin. Kinetic studies indicate that the increase in transcriptional potential is observed only when HMG-17 is incorporated into nucleosomes during chromatin assembly.

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Selected References

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

  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. Almouzni G., Clark D. J., Méchali M., Wolffe A. P. Chromatin assembly on replicating DNA in vitro. Nucleic Acids Res. 1990 Oct 11;18(19):5767–5774. doi: 10.1093/nar/18.19.5767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Almouzni G., Méchali M. Assembly of spaced chromatin involvement of ATP and DNA topoisomerase activity. EMBO J. 1988 Dec 20;7(13):4355–4365. doi: 10.1002/j.1460-2075.1988.tb03334.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Almouzni G., Méchali M., Wolffe A. P. Competition between transcription complex assembly and chromatin assembly on replicating DNA. EMBO J. 1990 Feb;9(2):573–582. doi: 10.1002/j.1460-2075.1990.tb08145.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Almouzni G., Méchali M., Wolffe A. P. Transcription complex disruption caused by a transition in chromatin structure. Mol Cell Biol. 1991 Feb;11(2):655–665. doi: 10.1128/mcb.11.2.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Almouzni G., Wolffe A. P. Nuclear assembly, structure, and function: the use of Xenopus in vitro systems. Exp Cell Res. 1993 Mar;205(1):1–15. doi: 10.1006/excr.1993.1051. [DOI] [PubMed] [Google Scholar]
  8. Banerjee S., Cantor C. R. Nucleosome assembly of simian virus 40 DNA in a mammalian cell extract. Mol Cell Biol. 1990 Jun;10(6):2863–2873. doi: 10.1128/mcb.10.6.2863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brawley J. V., Martinson H. G. HMG proteins 14 and 17 become cross-linked to the globular domain of histone H3 near the nucleosome core particle dyad. Biochemistry. 1992 Jan 21;31(2):364–370. doi: 10.1021/bi00117a008. [DOI] [PubMed] [Google Scholar]
  10. Bustin M., Becerra P. S., Crippa M. P., Lehn D. A., Pash J. M., Shiloach J. Recombinant human chromosomal proteins HMG-14 and HMG-17. Nucleic Acids Res. 1991 Jun 11;19(11):3115–3121. doi: 10.1093/nar/19.11.3115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Bustin M., Crippa M. P., Pash J. M. Immunochemical analysis of the exposure of high mobility group protein 14 and 17 surfaces in chromatin. J Biol Chem. 1990 Nov 25;265(33):20077–20080. [PubMed] [Google Scholar]
  12. Bustin M., Lehn D. A., Landsman D. Structural features of the HMG chromosomal proteins and their genes. Biochim Biophys Acta. 1990 Jul 30;1049(3):231–243. doi: 10.1016/0167-4781(90)90092-g. [DOI] [PubMed] [Google Scholar]
  13. Bustin M., Soares N., Landsman D., Srikantha T., Collins J. M. Cell cycle regulated synthesis of an abundant transcript for human chromosomal protein HMG-17. Nucleic Acids Res. 1987 Apr 24;15(8):3549–3561. doi: 10.1093/nar/15.8.3549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Camerini-Otero R. D., Sollner-Webb B., Felsenfeld G. The organization of histones and DNA in chromatin: evidence for an arginine-rich histone kernel. Cell. 1976 Jul;8(3):333–347. doi: 10.1016/0092-8674(76)90145-8. [DOI] [PubMed] [Google Scholar]
  15. Clark D. J., Felsenfeld G. A nucleosome core is transferred out of the path of a transcribing polymerase. Cell. 1992 Oct 2;71(1):11–22. doi: 10.1016/0092-8674(92)90262-b. [DOI] [PubMed] [Google Scholar]
  16. Crippa M. P., Alfonso P. J., Bustin M. Nucleosome core binding region of chromosomal protein HMG-17 acts as an independent functional domain. J Mol Biol. 1992 Nov 20;228(2):442–449. doi: 10.1016/0022-2836(92)90833-6. [DOI] [PubMed] [Google Scholar]
  17. Dilworth S. M., Black S. J., Laskey R. A. Two complexes that contain histones are required for nucleosome assembly in vitro: role of nucleoplasmin and N1 in Xenopus egg extracts. Cell. 1987 Dec 24;51(6):1009–1018. doi: 10.1016/0092-8674(87)90587-3. [DOI] [PubMed] [Google Scholar]
  18. Dong F., van Holde K. E. Nucleosome positioning is determined by the (H3-H4)2 tetramer. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10596–10600. doi: 10.1073/pnas.88.23.10596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Drew H. R. Reconstitution of short-spaced chromatin from the histone octamer and either HMG-14,17 or histone H1. J Mol Biol. 1993 Apr 5;230(3):824–836. doi: 10.1006/jmbi.1993.1204. [DOI] [PubMed] [Google Scholar]
  20. Druckmann S., Mendelson E., Landsman D., Bustin M. Immunofractionation of DNA sequences associated with HMG-17 in chromatin. Exp Cell Res. 1986 Oct;166(2):486–496. doi: 10.1016/0014-4827(86)90493-3. [DOI] [PubMed] [Google Scholar]
  21. Einck L., Bustin M. Inhibition of transcription in somatic cells by microinjection of antibodies to chromosomal proteins. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6735–6739. doi: 10.1073/pnas.80.22.6735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Einck L., Bustin M. The intracellular distribution and function of the high mobility group chromosomal proteins. Exp Cell Res. 1985 Feb;156(2):295–310. doi: 10.1016/0014-4827(85)90539-7. [DOI] [PubMed] [Google Scholar]
  23. Garel A., Axel R. Selective digestion of transcriptionally active ovalbumin genes from oviduct nuclei. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3966–3970. doi: 10.1073/pnas.73.11.3966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Glikin G. C., Ruberti I., Worcel A. Chromatin assembly in Xenopus oocytes: in vitro studies. Cell. 1984 May;37(1):33–41. doi: 10.1016/0092-8674(84)90298-8. [DOI] [PubMed] [Google Scholar]
  25. Grunstein M. Nucleosomes: regulators of transcription. Trends Genet. 1990 Dec;6(12):395–400. doi: 10.1016/0168-9525(90)90299-l. [DOI] [PubMed] [Google Scholar]
  26. Hansen J. C., Wolffe A. P. Influence of chromatin folding on transcription initiation and elongation by RNA polymerase III. Biochemistry. 1992 Sep 1;31(34):7977–7988. doi: 10.1021/bi00149a032. [DOI] [PubMed] [Google Scholar]
  27. Hayes J. J., Clark D. J., Wolffe A. P. Histone contributions to the structure of DNA in the nucleosome. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6829–6833. doi: 10.1073/pnas.88.15.6829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Hayes J. J., Wolffe A. P. The interaction of transcription factors with nucleosomal DNA. Bioessays. 1992 Sep;14(9):597–603. doi: 10.1002/bies.950140905. [DOI] [PubMed] [Google Scholar]
  29. Hebbes T. R., Thorne A. W., Crane-Robinson C. A direct link between core histone acetylation and transcriptionally active chromatin. EMBO J. 1988 May;7(5):1395–1402. doi: 10.1002/j.1460-2075.1988.tb02956.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Jackson V. In vivo studies on the dynamics of histone-DNA interaction: evidence for nucleosome dissolution during replication and transcription and a low level of dissolution independent of both. Biochemistry. 1990 Jan 23;29(3):719–731. doi: 10.1021/bi00455a019. [DOI] [PubMed] [Google Scholar]
  31. Kamakaka R. T., Thomas J. O. Chromatin structure of transcriptionally competent and repressed genes. EMBO J. 1990 Dec;9(12):3997–4006. doi: 10.1002/j.1460-2075.1990.tb07621.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kleinschmidt J. A., Scheer U., Dabauvalle M. C., Bustin M., Franke W. W. High mobility group proteins of amphibian oocytes: a large storage pool of a soluble high mobility group-1-like protein and involvement in transcriptional events. J Cell Biol. 1983 Sep;97(3):838–848. doi: 10.1083/jcb.97.3.838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kleinschmidt J. A., Steinbeisser H. DNA-dependent phosphorylation of histone H2A.X during nucleosome assembly in Xenopus laevis oocytes: involvement of protein phosphorylation in nucleosome spacing. EMBO J. 1991 Oct;10(10):3043–3050. doi: 10.1002/j.1460-2075.1991.tb07855.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Lam B. S., Carroll D. Tandemly repeated DNA sequences from Xenopus laevis. I. Studies on sequence organization and variation in satellite 1 DNA (741 base-pair repeat). J Mol Biol. 1983 Apr 25;165(4):567–585. doi: 10.1016/s0022-2836(83)80267-8. [DOI] [PubMed] [Google Scholar]
  36. Landsman D., Bustin M. Assessment of the transcriptional activation potential of the HMG chromosomal proteins. Mol Cell Biol. 1991 Sep;11(9):4483–4489. doi: 10.1128/mcb.11.9.4483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Landsman D., Mendelson E., Druckmann S., Bustin M. Exchange of proteins during immunofractionation of chromatin. Exp Cell Res. 1986 Mar;163(1):95–102. doi: 10.1016/0014-4827(86)90561-6. [DOI] [PubMed] [Google Scholar]
  38. Malik N., Smulson M., Bustin M. Enrichment of acetylated histones in polynucleosomes containing high mobility group protein 17 revealed by immunoaffinity chromatography. J Biol Chem. 1984 Jan 25;259(2):699–702. [PubMed] [Google Scholar]
  39. 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]
  40. McGhee J. D., Rau D. C., Felsenfeld G. The high mobility group proteins HMG 14 and 17, do not prevent the formation of chromatin higher order structure. Nucleic Acids Res. 1982 Mar 25;10(6):2007–2016. doi: 10.1093/nar/10.6.2007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Peterson R. C., Doering J. L., Brown D. D. Characterization of two xenopus somatic 5S DNAs and one minor oocyte-specific 5S DNA. Cell. 1980 May;20(1):131–141. doi: 10.1016/0092-8674(80)90241-x. [DOI] [PubMed] [Google Scholar]
  42. Piñeiro M., González P. J., Palacián E., Hernández F. Effect of high mobility group proteins 14 and 17 on the structural and transcriptional properties of acetylated complete and H2A.H2B-deficient nucleosomal cores. Arch Biochem Biophys. 1992 May 15;295(1):115–119. doi: 10.1016/0003-9861(92)90495-i. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Simpson R. T. Nucleosome positioning: occurrence, mechanisms, and functional consequences. Prog Nucleic Acid Res Mol Biol. 1991;40:143–184. doi: 10.1016/s0079-6603(08)60841-7. [DOI] [PubMed] [Google Scholar]
  45. Smith D. R., Jackson I. J., Brown D. D. Domains of the positive transcription factor specific for the Xenopus 5S RNA gene. Cell. 1984 Jun;37(2):645–652. doi: 10.1016/0092-8674(84)90396-9. [DOI] [PubMed] [Google Scholar]
  46. Tremethick D. J., Drew H. R. High mobility group proteins 14 and 17 can space nucleosomes in vitro. J Biol Chem. 1993 May 25;268(15):11389–11393. [PubMed] [Google Scholar]
  47. Tremethick D. J., Frommer M. Partial purification, from Xenopus laevis oocytes, of an ATP-dependent activity required for nucleosome spacing in vitro. J Biol Chem. 1992 Jul 25;267(21):15041–15048. [PubMed] [Google Scholar]
  48. 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]
  49. 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]
  50. 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]
  51. Westermann R., Grossbach U. Localization of nuclear proteins related to high mobility group protein 14 (HMG 14) in polytene chromosomes. Chromosoma. 1984;90(5):355–365. doi: 10.1007/BF00294162. [DOI] [PubMed] [Google Scholar]
  52. Wolffe A. P. Dominant and specific repression of Xenopus oocyte 5S RNA genes and satellite I DNA by histone H1. EMBO J. 1989 Feb;8(2):527–537. doi: 10.1002/j.1460-2075.1989.tb03407.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wolffe A. P. Implications of DNA replication for eukaryotic gene expression. J Cell Sci. 1991 Jun;99(Pt 2):201–206. doi: 10.1242/jcs.99.2.201. [DOI] [PubMed] [Google Scholar]
  54. Wolffe A. P., Morse R. H. The transcription complex of the Xenopus somatic 5 S RNA gene. A functional analysis of protein-DNA interactions outside of the internal control region. J Biol Chem. 1990 Mar 15;265(8):4592–4599. [PubMed] [Google Scholar]
  55. Wolffe A. P., Schild C. Chromatin assembly. Methods Cell Biol. 1991;36:541–559. [PubMed] [Google Scholar]
  56. Wolffe A. P. Transcription fraction TFIIIC can regulate differential Xenopus 5S RNA gene transcription in vitro. EMBO J. 1988 Apr;7(4):1071–1079. doi: 10.1002/j.1460-2075.1988.tb02915.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. van Holde K. E., Lohr D. E., Robert C. What happens to nucleosomes during transcription? J Biol Chem. 1992 Feb 15;267(5):2837–2840. [PubMed] [Google Scholar]

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