Surovtseva Yulia V, Shakirov Eugene V, Vespa Laurent, Osbun Nathan, Song Xiangyu, Shippen Dorothy E
Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843-2128, USA.
EMBO J. 2007 Aug 8;26(15):3653-61. doi: 10.1038/sj.emboj.7601792. Epub 2007 Jul 12.
POT1 is a single-copy gene in yeast and humans that encodes a single-strand telomere binding protein required for chromosome end protection and telomere length regulation. In contrast, Arabidopsis harbors multiple, divergent POT-like genes that bear signature N-terminal OB-fold motifs, but otherwise share limited sequence similarity. Here, we report that plants null for AtPOT1 show no telomere deprotection phenotype, but rather exhibit progressive loss of telomeric DNA. Genetic analysis indicates that AtPOT1 acts in the same pathway as telomerase. In vitro levels of telomerase activity in pot1 mutants are significantly reduced and are more variable than wild-type. Consistent with this observation, AtPOT1 physically associates with active telomerase particles. Although low levels of AtPOT1 can be detected at telomeres in unsynchronized cells and in cells arrested in G2, AtPOT1 binding is significantly enhanced during S-phase, when telomerase is thought to act at telomeres. Our findings indicate that AtPOT1 is a novel accessory factor for telomerase required for positive telomere length regulation, and they underscore the coordinate and extraordinarily rapid evolution of telomere proteins and the telomerase enzyme.
POT1在酵母和人类中是一个单拷贝基因,它编码一种染色体末端保护和端粒长度调控所需的单链端粒结合蛋白。相比之下,拟南芥含有多个不同的类POT基因,这些基因带有标志性的N端OB折叠基序,但在其他方面序列相似性有限。在这里,我们报告说,AtPOT1缺失的植物没有端粒去保护表型,而是表现出端粒DNA的逐渐丢失。遗传分析表明,AtPOT1与端粒酶作用于同一途径。pot1突变体中端粒酶活性的体外水平显著降低,且比野生型更具变异性。与这一观察结果一致,AtPOT1与活性端粒酶颗粒在物理上相互关联。尽管在未同步化的细胞和停滞在G2期的细胞的端粒中可以检测到低水平的AtPOT1,但在S期(据认为端粒酶在此时作用于端粒),AtPOT1的结合显著增强。我们的研究结果表明,AtPOT1是端粒酶的一种新型辅助因子,是正向端粒长度调控所必需的,并且强调了端粒蛋白和端粒酶的协同且极其快速的进化。