Virta-Pearlman V, Morris D K, Lundblad V
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
Genes Dev. 1996 Dec 15;10(24):3094-104. doi: 10.1101/gad.10.24.3094.
In Saccharomyces cerevisiae, deletion of the EST1 gene results in phenotypes identical to those displayed by a deletion of a known component of telomerase (the yeast telomerase RNA), arguing that EST1 is also critical for telomerase function. In this study, we show that the Estl protein binds to yeast G-rich telomeric oligonucleotides in vitro. Binding is specific for single-stranded substrates and requires a free 3' terminus, consistent with the properties expected for a protein bound to the 3' single-stranded G-rich extension present at the telomere. Assessment of the in vivo function of this single-stranded DNA-binding protein has shown that EST1 acts in the same pathway of telomere replication as the TLC1 telomerase RNA, by several different genetic criteria: est1 tlc1 double mutant strains show no enhancement of phenotype relative to either single mutant strain, and EST1 dominant mutations have an effect on telomeric silencing similar to that displayed by TLC1 previously. We propose that Est1 is a telomere end-binding protein that is required to mediate recognition of the end of the chromosome by telomerase.
在酿酒酵母中,EST1基因的缺失导致的表型与已知的端粒酶组分(酵母端粒酶RNA)缺失所表现出的表型相同,这表明EST1对端粒酶功能也至关重要。在本研究中,我们发现Estl蛋白在体外可与酵母富含G的端粒寡核苷酸结合。这种结合对单链底物具有特异性,并且需要一个游离的3'末端,这与预期与端粒处存在的3'单链富含G的延伸结合的蛋白质的特性一致。对这种单链DNA结合蛋白的体内功能评估表明,根据几种不同的遗传学标准,EST1与TLC1端粒酶RNA在端粒复制的同一途径中发挥作用:est1 tlc1双突变菌株相对于任一单突变菌株而言,其表型均未增强,并且EST1显性突变对端粒沉默的影响与之前TLC1所表现出的类似。我们提出,Est1是一种端粒末端结合蛋白,它是端粒酶介导识别染色体末端所必需的。