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Volume 270,
Number 38,
Issue of September 22, pp. 22514-22521, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Electrophoresis
of Chromatin on Nondenaturing Agarose Gels Containing Mg SELF-ASSEMBLY OF SMALL CHROMATIN FRAGMENTS AND FOLDING OF THE
30-nm FIBER
(Received for publication, May 15, 1995; and in revised form, June 30, 1995)
Salvador
Bartolomé,
Antonio
Bermúdez ,
Joan-Ramon
Daban
We show that nondenaturing agarose gels can be used for the
study of the structure and dynamic properties of native
(uncross-linked) chromatin. In gels containing 1.7 mM Mg , chicken erythrocyte chromatin fragments
having from about 6 to 50 nucleosomes produce well defined bands. These
bands have an electrophoretic mobility that decreases only slightly
with molecular weight. This surprising behavior is not observed in low
ionic strength gels. Fragments with less than 6 nucleosomes and low
content of histones H1-H5 give rise to broad bands in gels with
Mg . In contrast, fragments containing only 3-4
nucleosomes but with the normal H1-H5 content are able to form
associated structures with a mobility similar to that observed for high
molecular weight chromatin. Electron microscopy results indicate that
the associated fragments and the fragments of higher molecular weight
show similar electrophoretic properties because they become very
compact in the presence of Mg and form cylindrical
structures with a diameter of 33 nm. Our results suggest that the
interactions involved in the self-assembly of small fragments are the
same that direct the folding of larger fragments; in both cases, the
resulting compact chromatin structure is formed from a basic element
containing 5-7 nucleosomes.

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Copyright © 1995 by the American Society for Biochemistry and Molecular Biology.
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