Niida H, Matsumoto T, Satoh H, Shiwa M, Tokutake Y, Furuichi Y, Shinkai Y
Nippon Roche Research Center, Kamakura, Japan.
Nat Genet. 1998 Jun;19(2):203-6. doi: 10.1038/580.
The ribonucleoprotein enzyme telomerase synthesizes telomeric DNA onto chromosome ends. Telomere length is maintained, by the presence of telomerase activity, in the vast majority of primary tumours and stem cells, suggesting that telomere maintenance is essential for cellular immortalization. Recently, the telomerase RNA component in human and mouse (TERC and Terc, respectively), a telomerase-associated protein TEP1/TLP1 (refs 6,7) and the human catalytic subunit protein TERT (refs 8,9) have been identified. To examine the role of telomerase in telomere maintenance and cellular viability, we established Terc-deficient embryonic stem (ES) cells. It is known that telomerase activity is absent in cells from Terc-knockout mice. Although the study showed that telomere shortening was observed in the Terc-deficient cells from first to six generation animals, whether telomerase-dependent telomere maintenance was essential for cellular viability remained to be elucidated. To address this issue, we examined Terc-deficient ES cells under long-term culture conditions. Accompanying the continual telomere shortening, the growth rate of Terc-deficient ES cells was gradually reduced after more than 300 divisions. An impaired growth rate was maintained to approximately 450 divisions, and then cell growth virtually stopped. These data clearly show that telomerase-dependent telomere maintenance is critical for the growth of mammalian cells.
核糖核蛋白酶端粒酶将端粒DNA合成到染色体末端。在绝大多数原发性肿瘤和干细胞中,由于端粒酶活性的存在,端粒长度得以维持,这表明端粒维持对于细胞永生化至关重要。最近,已鉴定出人和小鼠中的端粒酶RNA组分(分别为TERC和Terc)、一种端粒酶相关蛋白TEP1/TLP1(参考文献6、7)以及人催化亚基蛋白TERT(参考文献8、9)。为了研究端粒酶在端粒维持和细胞活力中的作用,我们建立了Terc缺陷型胚胎干细胞(ES细胞)。已知Terc基因敲除小鼠的细胞中不存在端粒酶活性。尽管该研究表明在第一代至第六代动物的Terc缺陷型细胞中观察到了端粒缩短,但端粒酶依赖性端粒维持对于细胞活力是否必不可少仍有待阐明。为了解决这个问题,我们在长期培养条件下检测了Terc缺陷型ES细胞。随着端粒的持续缩短,Terc缺陷型ES细胞在超过300次分裂后生长速率逐渐降低。生长速率受损一直维持到大约450次分裂,然后细胞生长实际上停止了。这些数据清楚地表明,端粒酶依赖性端粒维持对于哺乳动物细胞的生长至关重要。