Smith C D, Smith D L, DeRisi J L, Blackburn E H
Department of Biochemistry and Biophysics, University of California, San Francisco, 94143-0448, USA.
Mol Biol Cell. 2003 Feb;14(2):556-70. doi: 10.1091/mbc.e02-08-0457.
In Saccharomyces cerevisiae, telomeric DNA is protected by a nonnucleosomal protein complex, tethered by the protein Rap1. Rif and Sir proteins, which interact with Rap1p, are thought to have further interactions with conventional nucleosomic chromatin to create a repressive structure that protects the chromosome end. We showed by microarray analysis that Rif1p association with the chromosome ends extends to subtelomeric regions many kilobases internal to the terminal telomeric repeats and correlates strongly with the previously determined genomic footprints of Rap1p and the Sir2-4 proteins in these regions. Although the end-protection function of telomeres is essential for genomic stability, telomeric DNA must also be copied by the conventional DNA replication machinery and replenished by telomerase, suggesting that transient remodeling of the telomeric chromatin might result in distinct protein complexes at different stages of the cell cycle. Using chromatin immunoprecipitation, we monitored the association of Rap1p, Rif1p, Rif2p, and the protein component of telomerase, Est2p, with telomeric DNA through the cell cycle. We provide evidence for dynamic remodeling of these components at telomeres.
在酿酒酵母中,端粒DNA由一种非核小体蛋白复合物保护,该复合物由Rap1蛋白连接。与Rap1p相互作用的Rif和Sir蛋白被认为与传统的核小体染色质有进一步的相互作用,以形成一种抑制性结构来保护染色体末端。我们通过微阵列分析表明,Rif1p与染色体末端的结合延伸到末端端粒重复序列内部许多千碱基的亚端粒区域,并且与这些区域中先前确定的Rap1p和Sir2 - 4蛋白的基因组足迹密切相关。虽然端粒的末端保护功能对于基因组稳定性至关重要,但端粒DNA也必须由传统的DNA复制机制进行复制,并由端粒酶进行补充,这表明端粒染色质的瞬时重塑可能导致细胞周期不同阶段出现不同的蛋白质复合物。利用染色质免疫沉淀技术,我们监测了Rap1p、Rif1p、Rif2p和端粒酶的蛋白质成分Est2p在细胞周期中与端粒DNA的结合情况。我们为这些成分在端粒处的动态重塑提供了证据。