Wang Bi-Dar, Eyre David, Basrai Munira, Lichten Michael, Strunnikov Alexander
Laboratory of Gene Regulation and Development, National Institute of Child Health and Human Development, National Cancer Institute, Bethesda, MD 20892, USA.
Mol Cell Biol. 2005 Aug;25(16):7216-25. doi: 10.1128/MCB.25.16.7216-7225.2005.
Mitotic chromosome condensation is chiefly driven by the condensin complex. The specific recognition (targeting) of chromosomal sites by condensin is an important component of its in vivo activity. We previously identified the rRNA gene cluster in Saccharomyces cerevisiae as an important condensin-binding site, but both genetic and cell biology data suggested that condensin also acts elsewhere. In order to characterize the genomic distribution of condensin-binding sites and to assess the specificity of condensin targeting, we analyzed condensin-bound sites using chromatin immunoprecipitation and hybridization to whole-genome microarrays. The genomic condensin-binding map shows preferential binding sites over the length of every chromosome. This analysis and quantitative PCR validation confirmed condensin-occupied sites across the genome and in the specialized chromatin regions: near centromeres and telomeres and in heterochromatic regions. Condensin sites were also enriched in the zones of converging DNA replication. Comparison of condensin binding in cells arrested in G(1) and mitosis revealed a cell cycle dependence of condensin binding at some sites. In mitotic cells, condensin was depleted at some sites while enriched at rRNA gene cluster, subtelomeric, and pericentromeric regions.
有丝分裂染色体凝聚主要由凝聚素复合物驱动。凝聚素对染色体位点的特异性识别(靶向作用)是其体内活性的重要组成部分。我们之前在酿酒酵母中鉴定出rRNA基因簇是一个重要的凝聚素结合位点,但遗传和细胞生物学数据均表明凝聚素在其他地方也发挥作用。为了描述凝聚素结合位点的基因组分布并评估凝聚素靶向作用的特异性,我们使用染色质免疫沉淀和全基因组微阵列杂交分析了凝聚素结合位点。基因组凝聚素结合图谱显示在每条染色体的全长上都有优先结合位点。该分析和定量PCR验证证实了全基因组以及在特殊染色质区域(着丝粒和端粒附近以及异染色质区域)存在凝聚素占据的位点。凝聚素位点在DNA复制汇合区域也有富集。比较处于G1期和有丝分裂期停滞的细胞中的凝聚素结合情况,发现凝聚素在某些位点的结合存在细胞周期依赖性。在有丝分裂细胞中,凝聚素在某些位点减少,而在rRNA基因簇、亚端粒和着丝粒周围区域富集。