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* Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA;
Department of Medicine, Biochemistry, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA;
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA
1Correspondence: Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA. E-mail: brinkley{at}bcm.tmc.edu
The centromere-kinetochore complex can be divided into distinct domains
based on structure and function. Previous work has used CREST
auto-antibodies with various microscopic techniques to map the
locations of proteins within the centromere-kinetochore complex and to
analyze the maturation of prekinetochores before mitosis. Here we have
focused on the centromere-specific histone Centromere Protein (CENP)-A
and its spatial relationship to other histones and histone
modifications found in condensed chromatin. We demonstrate that the
phosphorylation of histone H3 is essentially excluded from a specific
region of centromeric chromatin, defined by the presence of CENP-A.
Interspersion of CENP-B with phosphorylated H3 in the inner centromere
indicates that the exclusion of H3 modification is not a general
property of
-satellite DNA. We also demonstrate that these regions
are functionally distinct by fragmenting mitotic chromatin into motile
centromere-kinetochore fragments that contain CENP-A with little or no
phosphorylated H3 and nonmotile fragments that contain exclusively
phosphorylated H3. The sequence of CENP-A diverges from H3 in a number
of key residues involved in chromosome condensation and in
transcription, potentially allowing a more specialized chromatin
structure within centromeric heterochromatin, on which kinetochore
plates may nucleate and mature. This specialized centromere subdomain
would be predicted to have a very tight and static nucleosome structure
as a result of the absence of H3 phosphorylation and acetylation.Van
Hooser, A. A., Mancini, M. A., Allis, C. D., Sullivan, K. F., Brinkley,
B. R. The mammalian centromere: structural domains and the attenuation
of chromatin modeling.
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