Nucleic Acids Research, Vol 27, Issue 2 532-536, Copyright © 1999 by Oxford University Press
KA Bailey, CS Chow and JN Reeve
Recombinant (r)HMfB (archaealhistone B fromMethanothermusfervidus) formed
complexes with increasing stability with DNA molecules increasing in length
from 52 to 100 bp, but not with a 39 bp molecule. By using125I-labeled
rHMfB-YY (an rHMfB variant with I31Y and M35Y replacements) and32P-labeled
100 bp DNA, these complexes, designated archaeal nucleosomes, have been
shown to contain an archaeal histone tetramer. Consistent with DNA bending
and wrapping, addition of DNA ligase to archaeal nucleosomes assembled with
88 and 128 bp DNAs resulted in covalently-closed monomeric circular DNAs
which, following histone removal, were positively supercoiled based on
their electrophoretic mobilities in the presence of ethidium bromide before
and after relaxation by calf thymus topoisomerase I. Ligase addition to
mixtures of rHMfB with 53 or 30 bp DNA molecules also resulted in circular
DNAs but these were circular dimers and trimers. These short DNA molecules
apparently had to be ligated into longer linear multimers for assembly into
archaeal nucleosomes and ligation into circles. rHMfB assembled into
archaeal nucleosomes at lower histone to DNA ratios with the supercoiled,
circular ligation product than with the original 88 bp linear version of
this molecule. Archaeal histones are most similar to the globular histone
fold region of eukaryal histone H4, and the results reported are consistent
with archaeal nucleosomes resembling the structure formed by eukaryal
histone (H3+H4)2tetramers.
ARTICLES
Histone stoichiometry and DNA circularization in archaeal nucleosomes
Department of Microbiology, Ohio State University, Columbus, OH 43210, USA and Department of Chemistry,Wayne State University, Detroit, MI 48202, USA.
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