Webb Christopher J, Zakian Virginia A
Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
Nat Struct Mol Biol. 2008 Jan;15(1):34-42. doi: 10.1038/nsmb1354. Epub 2007 Dec 23.
Although the catalytic subunit of the Schizosaccharomyces pombe telomerase holoenzyme was identified over ten years ago, the unusual heterogeneity of its telomeric DNA made it difficult to identify its RNA component. We used a new two-step immunoprecipitation and reverse transcription-PCR technique to identify the S. pombe telomerase RNA, which we call TER1. TER1 RNA was 1,213 nucleotides long, similar in size to the Saccharomyces cerevisiae telomerase RNA, TLC1. TER1 RNA associated in vivo with the two known subunits of the S. pombe telomerase holoenzyme, Est1p and Trt1p, and neither association was dependent on the other holoenzyme component. We present a model to explain how telomerase introduces heterogeneity into S. pombe telomeres. The technique used here to identify TER1 should be generally applicable to other model organisms.
尽管粟酒裂殖酵母端粒酶全酶的催化亚基早在十多年前就已被鉴定出来，但其端粒DNA异常的异质性使得鉴定其RNA组分变得困难。我们使用了一种新的两步免疫沉淀和逆转录PCR技术来鉴定粟酒裂殖酵母端粒酶RNA，我们将其命名为TER1。TER1 RNA长1213个核苷酸，大小与酿酒酵母端粒酶RNA TLC1相似。TER1 RNA在体内与粟酒裂殖酵母端粒酶全酶的两个已知亚基Est1p和Trt1p相关联，且这两种关联均不依赖于其他全酶组分。我们提出了一个模型来解释端粒酶如何将异质性引入粟酒裂殖酵母的端粒中。这里用于鉴定TER1的技术应该普遍适用于其他模式生物。
