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A more recent version of this article appeared on September 1, 2003
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Submitted on January 8, 2003
Accepted on April 19, 2003
1 Institute of Molecular Biology and Pathology, CNR, c/o Dept. of Genetics and Molecular Biology, University "La Sapienza", 00185 Rome (Italy); Department of Biology, CB#3280, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599 (USA)
2 Institute of Molecular Biology and Pathology, CNR, c/o Dept. of Genetics and Molecular Biology, University "La Sapienza", 00185 Rome (Italy)
* Corresponding author. E-mail address: f.degrassi{at}caspur.it.
Posttranslational modifications of core histones give a major
contribution in driving changes in chromatin conformation and
compaction. Here we investigated the role of histone deacetylation on
the mitotic process by inhibiting histone deacetylases shortly before
mitosis in human primary fibroblasts. Cells entering mitosis with
hyperacetylated histones displayed altered chromatin conformation
associated with decreased reactivity to the anti Ser 10 phospho H3
antibody, increased recruitment of protein phosphatase 1-
on mitotic
chromosomes, and depletion of heterochromatin protein 1 from the
centromeric heterochromatin. Inhibition of histone deacetylation
before mitosis produced defective chromosome condensation and
impaired mitotic progression in living cells, suggesting that improper
chromosome condensation may induce mitotic checkpoint activation. In
situ hybridization analysis on anaphase cells demonstrated the presence
of chromatin bridges, which were caused by persisting cohesion along
sister chromatid arms after centromere separation. Thus, the presence
of hyperacetylated chromatin during mitosis impairs proper chromosome
condensation during the preanaphase stages, resulting in poor sister
chromatid resolution. Lagging chromosomes consisting of single or
paired sisters were also induced by the presence of hyperacetylated
histones, indicating that the less constrained centromeric organization
associated with heterochromatin protein 1 depletion may promote the
attachment of kinetochores to microtubules coming from both
poles.
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