Combined Program on Microbiology and Immunology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, 565-0871 Osaka, Japan.
Sci Rep. 2013 Nov 8;3:3171. doi: 10.1038/srep03171.
Biochemical analysis of molecular interactions in specific genomic regions requires their isolation while retaining molecular interactions in vivo. Here, we report isolation of telomeres by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using a transcription activator-like (TAL) protein recognizing telomere repeats. Telomeres recognized by the tagged TAL protein were immunoprecipitated with an antibody against the tag and subjected to identification of telomere-binding molecules. enChIP-mass spectrometry (enChIP-MS) targeting telomeres identified known and novel telomere-binding proteins. The data have been deposited to the ProteomeXchange with identifier PXD000461. In addition, we showed that RNA associated with telomeres could be isolated by enChIP. Identified telomere-binding molecules may play important roles in telomere biology. enChIP using TAL proteins would be a useful tool for biochemical analysis of specific genomic regions of interest.
特定基因组区域中分子相互作用的生化分析需要在保留体内分子相互作用的情况下对其进行分离。在这里，我们报告了使用识别端粒重复序列的转录激活样（TAL）蛋白工程化 DNA 结合分子介导的染色质免疫沉淀（enChIP）来分离端粒。用针对标记的抗体将标记的 TAL 蛋白识别的端粒进行免疫沉淀，并对端粒结合分子进行鉴定。针对端粒的 enChIP-质谱（enChIP-MS）鉴定了已知和新的端粒结合蛋白。该数据已被 ProteomeXchange 以标识符 PXD000461 存储。此外，我们还表明，通过 enChIP 可以分离与端粒相关的 RNA。鉴定出的端粒结合分子可能在端粒生物学中发挥重要作用。使用 TAL 蛋白的 enChIP 将成为对特定感兴趣基因组区域进行生化分析的有用工具。
