Nucleosomal arrays can be salt-reconstituted on a single-copy MMTV promoter DNA template: Their properties differ in several ways from those of comparable 5S concatameric arrays

R. Bash, H. Wang, J. Yodh, G. Hager, Stuart Lindsay, D. Lohr

Research output: Contribution to journalArticle

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Abstract

Subsaturated nucleosomal arrays were reconstituted on a single-copy MMTV promoter DNA fragment by salt dialysis procedures and studied by atomic force microscopy. Up to an occupation level of approximately eight nucleosomes on this 1900 bp template, salt reconstitution produces nucleosomal arrays which look very similar to comparably loaded 5S rDNA nucleosomal arrays; i.e., nucleosomes are dispersed on the DNA template. Thus, at these occupation levels, the single-copy MMTV template forms arrays suitable for biophysical analyses. A quantitative comparison of the population features of subsaturated MMTV and 5S arrays detects differences between the two: a requirement for higher histone levels to achieve a given level of nucleosome occupation on MMTV templates, indicating that nucleosome loading is thermodynamically less favorable on this template; a preference for pairwise nucleosome occupation of the MMTV (but not the 5S) template at midrange occupation levels; and an enhanced salt stability for nucleosomes on MMTV versus 5S arrays, particularly in the midrange of array occupation. When average occupation levels exceed approximately eight nucleosomes per template, MMTV arrays show a significant level of mainly intramolecular compaction; 5S arrays do not. Taken together, these results show clearly that the nature of the underlying DNA template can affect the physical properties of nucleosomal arrays. DNA sequence-directed differences in the physical properties of chromatin may have important consequences for functional processes such as gene regulation.

Original languageEnglish (US)
Pages (from-to)4681-4690
Number of pages10
JournalBiochemistry
Volume42
Issue number16
DOIs
StatePublished - Apr 29 2003

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Nucleosomes
Occupations
Salts
DNA
Physical properties
Dialysis
Atomic Force Microscopy
DNA sequences
Ribosomal DNA
Gene expression
Histones
Chromatin
Atomic force microscopy
Compaction
Population
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nucleosomal arrays can be salt-reconstituted on a single-copy MMTV promoter DNA template : Their properties differ in several ways from those of comparable 5S concatameric arrays. / Bash, R.; Wang, H.; Yodh, J.; Hager, G.; Lindsay, Stuart; Lohr, D.

In: Biochemistry, Vol. 42, No. 16, 29.04.2003, p. 4681-4690.

Research output: Contribution to journalArticle

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abstract = "Subsaturated nucleosomal arrays were reconstituted on a single-copy MMTV promoter DNA fragment by salt dialysis procedures and studied by atomic force microscopy. Up to an occupation level of approximately eight nucleosomes on this 1900 bp template, salt reconstitution produces nucleosomal arrays which look very similar to comparably loaded 5S rDNA nucleosomal arrays; i.e., nucleosomes are dispersed on the DNA template. Thus, at these occupation levels, the single-copy MMTV template forms arrays suitable for biophysical analyses. A quantitative comparison of the population features of subsaturated MMTV and 5S arrays detects differences between the two: a requirement for higher histone levels to achieve a given level of nucleosome occupation on MMTV templates, indicating that nucleosome loading is thermodynamically less favorable on this template; a preference for pairwise nucleosome occupation of the MMTV (but not the 5S) template at midrange occupation levels; and an enhanced salt stability for nucleosomes on MMTV versus 5S arrays, particularly in the midrange of array occupation. When average occupation levels exceed approximately eight nucleosomes per template, MMTV arrays show a significant level of mainly intramolecular compaction; 5S arrays do not. Taken together, these results show clearly that the nature of the underlying DNA template can affect the physical properties of nucleosomal arrays. DNA sequence-directed differences in the physical properties of chromatin may have important consequences for functional processes such as gene regulation.",
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