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Departments of Anatomy and Medical Biochemistry, Faculty of
Medicine, University of Calgary, Calgary, Alberta, Canada T2N 4N1
Histones found within transcriptionally competent and active
regions of the genome are highly acetylated. Moreover, these highly
acetylated histones have very short half-lives. Thus, both histone
acetyltransferases and histone deacetylases must enrich within or near
these euchromatic regions of the interphase chromatids. Using an
antibody specific for highly acetylated histone H3, we have
investigated the organization of transcriptionally active and competent
chromatin as well as nuclear histone acetyltransferase and deacetylase
activities. We observe an exclusion of highly acetylated chromatin
around the periphery of the nucleus and an enrichment near
interchromatin granule clusters (IGCs). The highly acetylated chromatin
is found in foci that may reflect the organization of highly acetylated
chromatin into "chromonema" fibers. Transmission electron
microscopy of Indian muntjac fibroblast cell nuclei indicates that the
chromatin associated with the periphery of IGCs remains relatively
condensed, most commonly found in domains containing chromatin folded
beyond 30 nm. Using electron spectroscopic imaging, we demonstrate that
IGCs are clusters of ribonucleoprotein particles. The individual
granules comprise RNA-rich fibrils or globular regions that fold into
individual granules. Quantitative analysis of individual granules
indicates that they contain variable amounts of RNA estimated between
1.5 and >10 kb. We propose that interchromatin granules are
heterogeneous nuclear RNA-containing particles, some of which may be
pre-mRNA generated by nearby transcribed chromatin. An intermediary
zone between the IGC and surrounding chromatin is described that
contains factors with the potential to provide specificity to the
localization of sequences near IGCs.
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