Miller M C, Collins K
Department of Molecular and Cell Biology, University of California, Berkeley, 94720, USA.
Mol Cell. 2000 Oct;6(4):827-37. doi: 10.1016/s1097-2765(05)00078-x.
The telomerase enzyme adds simple sequence repeats to chromosome ends. Telomerases share two essential subunits, telomerase RNA and telomerase reverse transcriptase, that associate with species-specific proteins of predominantly unknown functions. The Tetrahymena p80/p95 complex can coimmunopurify active telomerase from cell extract, and recombinant p80/p95 can interact directly with telomerase RNA and single-stranded telomeric DNA in vitro. Here, we test the functions of p80/p95 in vivo. Surprisingly, telomerase RNA accumulation and telomerase activity in cell extract are unaffected by loss of the genes encoding p80/p95. However, in the absence of p80/p95, telomeres become elongated in both macronuclei and micronuclei. Micronuclear chromosome maintenance is also compromised. These findings suggest that p80/p95 functions to maintain appropriate telomere length and micronuclear genomic stability but does so in a manner different than previously anticipated.
端粒酶可将简单序列重复序列添加到染色体末端。端粒酶共有两个必需亚基,即端粒酶RNA和端粒酶逆转录酶,它们与功能大多未知的物种特异性蛋白质相关联。嗜热四膜虫的p80/p95复合物能够从细胞提取物中通过共免疫纯化获得活性端粒酶,并且重组p80/p95在体外可直接与端粒酶RNA和单链端粒DNA相互作用。在此,我们在体内测试了p80/p95的功能。令人惊讶的是,细胞提取物中端粒酶RNA的积累和端粒酶活性不受编码p80/p95的基因缺失的影响。然而,在缺乏p80/p95的情况下,大核和微核中的端粒都会变长。微核染色体的维持也会受到损害。这些发现表明,p80/p95的功能是维持适当的端粒长度和微核基因组稳定性,但其方式与先前预期的不同。