In order to better characterize linker histone interactions with DNA, avian erythrocyte-specific linker H5 and the trypsin-resistant globular domain of H5 (GH5) were used in DNA binding studies. To begin, H5 displayed a considerably higher binding
affinity for DNA than the isolated globular domain (GH5), supporting the importance of the...
Understanding the structure of the 30 nm fiber in chromatin is relevant to understanding eukaryotic replication and transcription. The major controversy among the models of the fiber concerns the disposition of the linker DNA, the DNA between adjacent nucleosomes, and the location of the linker histones. To determine if the...
The aim of this study was to gain more information about the
interactions between DNA and the histone octamer during the
process of transcription. This work used a pUC8 plasmid derivative
that contained the core promoter region of the RNA polymerase I of
Acanthamoeba castellanii, placed upstream of four repeats...
Time-resolved fluorescence spectroscopy was used to monitor the effects of
varying ionic strength on nucleosome core particle structure. Two main methods were
used in these studies. First, the fluorescence anisotropy decay of bound ethidium was
measured and was shown to reflect the rotational tumbling of the core particle through
solution,...
By using immobilized trypsin, I have been able to
prepare well-defined, stable trypsinized nucleosomes. The
difficulties of lacking of control in the extent of
trypsinization, which were encountered in previous studies
with the use of free trypsin, have been eliminated. The
nucleosomes and oligonucleosomes prepared by immobilized
trypsin are suitable...
Five forms of chromatin, which represent the in vivo folded forms
of nucleic acid, were isolated and used as the binding substrate for
model intercalating compound, ethidium bromide. For all forms of
chromatin, the affinity, location and structural effects of binding
were examined. On the level of the nucleosome, the...
Several levels of eukaryotic chromatin structure have
been observed: the nucleosome, the 10 nm and 30 nm fibers
and loop domains, apparently attached to the nuclear
matrix. In this research, the structure and function of
chromatin at two of these levels was investigated, with
studies on both nucleosome positioning and...
I examined, in three separate studies, the active chromatin
structure of the yeast, Saccharomyces cerevisiae.
Yeast contains four proteins having amino acid compositions
typical of the high mobility group (HMG) proteins. Three of these
are eluted from chromatin by 0.35 M NaCl; one is not, but is eluted
by 0.25...
The structure and transcriptional activity of intranuclear
and isolated chromatin from logarithmically growing
yeast cells has been compared to chromatin from cells which
have entered the stationary phase and ceased growing. Both
chromatins show a similar nucleosomal repeat pattern and a
160 by repeat size when digested with staphlococcal nuclease....
The nucleosome appears to be the basic structural unit of the
eukaryotic chromosome. The usual nucleosome preparation, isolated
from a micrococcal nuclease digest of nuclei, is heterogenous in DNA
length and protein composition with some particles containing spacer
DNA and associated protein. This heterogeneity complicated physical
and reconstitution studies. As...