• Molecules and Cells
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• v.38 no.9
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• pp.789-795
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• 2015

A chromosome territory is composed of chromosomal subdomains. The internal structure of chromosomal subdomains provides a structural framework for many genomic activities such as replication and DNA repair, and thus is key to determining the basis of their mechanisms. However, the internal structure and regulating proteins of a chromosomal subdomain remains elusive. Previously, we showed that the chromosome territory expanded after BAF53 knockdown. Because the integrity of chromosomal subdomains is a deciding factor of the volume of a chromosome territory, we examined here the effect of BAF53 knockdown on chromosomal subdomains. We found that BAF53 knockdown led to the disintegration of histone H2B-GFP-visualized chromosomal subdomains and BrdU-labeled replication foci. In addition, the size of DNA loops measured by the maximum fluorescent halo technique increased and became irregular after BAF53 knockdown, indicating DNA loops were released from the residual nuclear structure. These data can be accounted for by the model that BAF53 is prerequisite for maintaining the structural integrity of chromosomal subdomains.

• Animal cells and systems
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• v.16 no.3
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• pp.183-189
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• 2012

It has been suggested that chromatin is organized into the stable structures that provide fundamental units of chromosome architecture in interphase mammalian cells. The stable structures of chromatin can be visualized as replication foci when replicating DNA is labeled with thymidine analogs. Previously, we showed that the chromosome territory expanded after BAF53 knockdown. In this study, we found that BAF53 is required for the formation of replication foci. DNA replication was not impaired in BAF53 knockdown cells, suggesting that the decrease in the number of replication foci is due to disintegration of replication foci, but not suppression of DNA replication. The attractive forces that maintain structural integrity of replication foci could be disrupted by BAF53 knockdown, and it may be responsible, at least in part, for the expansion of chromosome territories after BAF53 knockdown.

• Animal cells and systems
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• v.13 no.4
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• pp.405-409
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• 2009

When DNA double-strand breaks (DSBs) were induced in mammalian cells, many DNA damage response proteins are accumulated at damage sites to form nuclear foci called IR-induced foci. Although the formation of foci has been shown to promote repair efficiency, the structural organization of chromatin in foci remains obscure. BAF53 is an actin-related protein which is required for maintenance of chromosome territory. In this study, we show that the formation of IR-induced foci by $\gamma$-H2AX and 53BP1 were reduced when BAF53 is depleted, while DSB- activated ATM pathway and the phosphorylation of H2AX remains intact after DNA damage in BAF53 knockdown cells. We also found that DSB repair efficiency was largely compromised in BAF53 knockdown cells. These results indicate that BAF53 is critical for formation of foci by $\gamma$-H2AX decorated chromatin at damage sites and the structural organization of chromatin in foci is an important factor to achieve the maximum efficiency of DNA repair.