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PMID: 11114182 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Fast kinetics of chromatin assembly revealed by single-molecule videomicroscopy and scanning force microscopy.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 26 ·2000-12-19 ·Pages 14251-6

Ladoux B, Quivy JP, Doyle P, du Roure O, Almouzni G, Viovy JL

Abstract

Fluorescence videomicroscopy and scanning force microscopy were used to follow, in real time, chromatin assembly on individual DNA molecules immersed in cell-free systems competent for physiological chromatin assembly. Within a few seconds, molecules are already compacted into a form exhibiting strong similarities to native chromatin fibers. In these extracts, the compaction rate is more than 100 times faster than expected from standard biochemical assays. Our data provide definite information on the forces involved (a few piconewtons) and on the reaction path. DNA compaction as a function of time revealed unique features of the assembly reaction in these extracts. They imply a sequential process with at least three steps, involving DNA wrapping as the final event. An absolute and quantitative measure of the kinetic parameters of the early steps in chromatin assembly under physiological conditions could thus be obtained.

MeSH Terms
Animals Bacteriophage lambda/genetics Cell Extracts Chromatin/metabolism DNA, Viral/metabolism Drosophila Kinetics Microscopy, Atomic Force/methods Microscopy, Video/methods Models, Molecular Nucleosomes/metabolism Time Factors Xenopus
Chemicals
Cell Extracts Chromatin DNA, Viral Nucleosomes
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ladoux B
Laboratoire de Physico-Chimie Curie (Unité Mixte de Recherche Centre National de la Recherche Scientifique/Institut Curie 168), Paris Cedex, France.
Quivy J P
Doyle P
du Roure O
Almouzni G
Viovy J L
References (37)
37 references, click to expand
  1. Protamine-induced condensation and decondensation of the same DNA molecule.
    Science. 1999 Oct 1;286(5437):120-3 PMID: 10506559
  2. Three-dimensional structure of extended chromatin fibers as revealed by tapping-mode scanning force microscopy.
    Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11621-5 PMID: 7972114
  3. Single-molecule studies of the effect of template tension on T7 DNA polymerase activity.
    Nature. 2000 Mar 2;404(6773):103-6 PMID: 10716452
  4. Single-molecule study of transcriptional pausing and arrest by E. coli RNA polymerase.
    Science. 2000 Mar 31;287(5462):2497-500 PMID: 10741971
  5. Dynamics of a tethered polymer in shear flow.
    Phys Rev Lett. 2000 May 15;84(20):4769-72 PMID: 10990792
  6. Assembly of SV40 chromatin in a cell-free system from Xenopus eggs.
    Cell. 1977 Feb;10(2):237-43 PMID: 189936
  7. Preferential association of newly synthesized histones with replicating SV40 DNA.
    Cell. 1977 Dec;12(4):947-51 PMID: 202394
  8. Preferential association of newly synthesized H3 and H4 histones with newly replicated DNA.
    J Biochem. 1978 Oct;84(4):985-8 PMID: 711710
  9. Assembly of newly replicated chromatin.
    Cell. 1978 Nov;15(3):969-77 PMID: 103629
  10. A new procedure for purifying histone pairs H2A + H2B and H3 + H4 from chromatin using hydroxylapatite.
    Nucleic Acids Res. 1979 Feb;6(2):689-96 PMID: 424310
  11. DNA folding by histones: the kinetics of chromatin core particle reassembly and the interaction of nucleosomes with histones.
    J Mol Biol. 1979 May 15;130(2):103-34 PMID: 469938
  12. Acidic polypeptides can assemble both histones and chromatin in vitro at physiological ionic strength.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):5000-4 PMID: 291918
  13. Role of histone pairs H2A,H2B and H3,H4 in the self-assembly of nucleosome core particles.
    J Mol Biol. 1982 Apr 25;156(4):771-89 PMID: 7120393
  14. Binding of ethidium bromide causes dissociation of the nucleosome core particle.
    Proc Natl Acad Sci U S A. 1986 Nov;83(22):8472-6 PMID: 3464964
  15. Deposition of newly synthesized histones: new histones H2A and H2B do not deposit in the same nucleosome with new histones H3 and H4.
    Biochemistry. 1987 Apr 21;26(8):2315-25 PMID: 3620448
  16. In vitro transcription of the Drosophila engrailed gene.
    Genes Dev. 1988 Jan;2(1):68-81 PMID: 3356339
  17. Deposition of newly synthesized histones: hybrid nucleosomes are not tandemly arranged on daughter DNA strands.
    Biochemistry. 1988 Mar 22;27(6):2109-20 PMID: 3378048
  18. Assembly of spaced chromatin promoted by DNA synthesis in extracts from Xenopus eggs.
    EMBO J. 1988 Mar;7(3):665-72 PMID: 3396538
  19. Expression of a histone H1-like protein is restricted to early Xenopus development.
    Genes Dev. 1988 Oct;2(10):1284-95 PMID: 3060404
  20. Assembly of spaced chromatin involvement of ATP and DNA topoisomerase activity.
    EMBO J. 1988 Dec 20;7(13):4355-65 PMID: 2854062
  21. Assembly and disassembly of the Drosophila RNA polymerase II complex during transcription.
    J Biol Chem. 1990 Feb 15;265(5):2624-31 PMID: 1689291
  22. 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):719-31 PMID: 1692479
  23. Chromatin assembly on replicating DNA in vitro.
    Nucleic Acids Res. 1990 Oct 11;18(19):5767-74 PMID: 2216769
  24. The mechanism of nucleosome assembly onto oligomers of the sea urchin 5 S DNA positioning sequence.
    J Biol Chem. 1991 Mar 5;266(7):4276-82 PMID: 1900288
  25. Role of linker histones in extended chromatin fibre structure.
    Nat Struct Biol. 1994 Nov;1(11):761-3 PMID: 7634084
  26. Chromatin unfolds.
    Cell. 1996 Jul 12;86(1):13-9 PMID: 8689680
  27. Single polymer dynamics in an elongational flow.
    Science. 1997 Jun 27;276(5321):2016-21 PMID: 9197259
  28. Crystal structure of the nucleosome core particle at 2.8 A resolution.
    Nature. 1997 Sep 18;389(6648):251-60 PMID: 9305837
  29. Alteration of nucleosome structure as a mechanism of transcriptional regulation.
    Annu Rev Biochem. 1998;67:545-79 PMID: 9759497
  30. Energy landscapes of receptor-ligand bonds explored with dynamic force spectroscopy.
    Nature. 1999 Jan 7;397(6714):50-3 PMID: 9892352
  31. Twenty-five years of the nucleosome, fundamental particle of the eukaryote chromosome.
    Cell. 1999 Aug 6;98(3):285-94 PMID: 10458604
  32. Stepwise assembly of chromatin during DNA replication in vitro.
    EMBO J. 1991 Apr;10(4):971-80 PMID: 1849080
  33. Cell-free system for assembly of transcriptionally repressed chromatin from Drosophila embryos.
    Mol Cell Biol. 1992 May;12(5):2241-9 PMID: 1569951
  34. Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads.
    Science. 1992 Nov 13;258(5085):1122-6 PMID: 1439819
  35. Disruption of the nucleosomes at the replication fork.
    EMBO J. 1993 Dec;12(12):4533-45 PMID: 8223463
  36. Relaxation of a single DNA molecule observed by optical microscopy.
    Science. 1994 May 6;264(5160):822-6 PMID: 8171336
  37. Pulling a single chromatin fiber reveals the forces that maintain its higher-order structure.
    Proc Natl Acad Sci U S A. 2000 Jan 4;97(1):127-32 PMID: 10618382
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.250471597
Published
2000-12-19
Pages
14251-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC18904
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.250471597 PMC Full Text View journal details
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