Liu Z P, Tye B K
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Genes Dev. 1991 Jan;5(1):49-59. doi: 10.1101/gad.5.1.49.
We have identified three yeast proteins that bind to poly(C.A)/poly(T.G) repeats characteristic of telomeric sequences from yeast to human. TBF alpha binds to the telomeric sequences of yeast, Tetrahymena, and vertebrates. In contrast, TBF beta binds only to yeast telomeric sequences. Also identified was RAP1, the transcriptional silencer protein, which binds to a sequence motif found in upstream activating sequences (UASs) of a number of genes; the sequence motif also occurs frequently in yeast telomeric sequences. Because poly(C.A)/poly(T.G) sequences from a wide range of organisms will serve as the primer for the in vivo extension of telomeres in yeast, TBF alpha is of particular interest. DNase I footprinting analysis indicated that TBF alpha binds to the junction between the subtelomeric X sequence and poly(C1-3A) in a cloned yeast telomere. Examination of the junctions of known X sequences indicated that they all contain one or more repeats of CCCTAA, a sequence that is repeated in vertebrate telomeres. Earlier, Murray et al. (1988) reported that heterologous telomeric sequences positioned as far as several hundred base pairs from the termini of linear molecules can allow the addition of yeast telomeric sequences from nontelomeric termini in vivo. A possible function for TBF alpha might be to serve as an anchoring protein for the yeast telomerase by binding to the conserved junction sequence at a distance from the terminus to allow addition of an irregular repeating sequence at the chromosome end.
我们已经鉴定出三种酵母蛋白,它们能与从酵母到人类的端粒序列所特有的聚(C.A)/聚(T.G)重复序列结合。TBFα能与酵母、四膜虫和脊椎动物的端粒序列结合。相比之下,TBFβ仅与酵母端粒序列结合。我们还鉴定出了RAP1,即转录沉默蛋白,它能与许多基因的上游激活序列(UASs)中发现的一个序列基序结合;该序列基序在酵母端粒序列中也经常出现。由于来自多种生物的聚(C.A)/聚(T.G)序列可作为酵母中端粒体内延伸的引物,因此TBFα特别令人感兴趣。DNA酶I足迹分析表明,TBFα与克隆的酵母端粒中近端粒X序列和聚(C1 - 3A)之间的连接处结合。对已知X序列连接处的检查表明,它们都包含一个或多个CCCTAA重复序列,该序列在脊椎动物端粒中也有重复。早些时候,默里等人(1988年)报道,位于线性分子末端数百个碱基对之外的异源端粒序列可在体内允许从非端粒末端添加酵母端粒序列。TBFα的一个可能功能可能是通过与距末端一定距离的保守连接序列结合,作为酵母端粒酶的锚定蛋白,从而允许在染色体末端添加不规则的重复序列。