酵母端粒长度调节因子TEL2编码一种与端粒DNA结合的蛋白质。
The yeast telomere length regulator TEL2 encodes a protein that binds to telomeric DNA.
作者信息
Kota R S, Runge K W
机构信息
The Lerner Research Institute, Cleveland Clinic Foundation, Department of Molecular Biology, NC 20, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
出版信息
Nucleic Acids Res. 1998 Mar 15;26(6):1528-35. doi: 10.1093/nar/26.6.1528.
TEL2 is required for telomere length regulation and viability in Saccharomyces cerevisiae. To investigate the mechanism by which Tel2p regulates telomere length, the majority (65%) of the TEL2 ORF was fused to the 3'-end of the gene for maltose binding protein, expressed in bacteria and the purified protein used in DNA binding studies. Rap1p, the major yeast telomere binding protein, recognizes a 13 bp duplex site 5'-GGTGTGTGGGTGT-3' in yeast telomeric DNA with high affinity. Gel shift experiments revealed that the MBP-Tel2p fusion binds the double-stranded yeast telomeric Rap1p site in a sequence-specific manner. Analysis of mutated sites showed that MBP-Tel2p could bind 5'-GTGTGTGG-3' within this 13 bp site. Methylation interference analysis revealed that Tel2p contacts the 5'-terminal guanine in the major groove. MBP-Tel2p did not bind duplex telomeric DNA repeats from vertebrates, Tetrahymena or Oxytricha. These results suggest that Tel2p is a DNA binding protein that recognizes yeast telomeric DNA.
端粒长度调节和酿酒酵母生存能力需要TEL2。为了研究Tel2p调节端粒长度的机制，将大部分（65%）的TEL2开放阅读框与麦芽糖结合蛋白基因的3'端融合，在细菌中表达，并将纯化的蛋白用于DNA结合研究。Rap1p是主要的酵母端粒结合蛋白，以高亲和力识别酵母端粒DNA中的13 bp双链位点5'-GGTGTGTGGGTGT-3'。凝胶迁移实验表明，MBP-Tel2p融合蛋白以序列特异性方式结合双链酵母端粒Rap1p位点。对突变位点的分析表明，MBP-Tel2p可以结合该13 bp位点内的5'-GTGTGTGG-3'。甲基化干扰分析表明，Tel2p在大沟中与5'末端鸟嘌呤接触。MBP-Tel2p不结合来自脊椎动物、四膜虫或游仆虫的双链端粒DNA重复序列。这些结果表明，Tel2p是一种识别酵母端粒DNA的DNA结合蛋白。
Zotero 插件