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DNA-Binding properties of the replication telomere protein

机译：复制端粒蛋白的DNA结合特性

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The replication Telomere Protein, rTP, is a nuclear protein from the ciliate Euplotes crassus that appears to be a novel telomere replication factor. rTP shares extensive amino acid sequence identity with the two proteins that bind and protect the macronuclear telomeres from the ciliates Oxytricha and Euplotes. Since the most extended regions of conservation fall within the DNA-binding domains of the telomere-binding proteins, when rTP was first identified it was predicted to be another structural telomere-binding protein. However, subsequent research demonstrated that rTP transcripts accumulate only during DNA replication and the rTP protein localizes to the sites of DNA replication within Euplotes macronuclei. We have now expressed rTP in a heterologous expression system and have examined the DNA-binding properties of the recombinant protein. We show that rTP binds specifically to the G-strand of Euplotes telomeric DNA and hence has some of the same DNA-binding characteristics as the Euplotes and Oxytricha telomere-binding proteins. However, other aspects of rTP binding are unique. In particular, the protein exhibits a very high off-rate and can bind double-stranded DNA as well as internal tracts of telomeric sequence. We conclude that rTP and the telomere-binding proteins are members of a class of proteins that have a conserved DNA-binding motif tailored to bind the G-strand of telomeric DNA. However, the unique DNA-binding characteristics of rTP indicate that the protein has evolved to fulfil a specialized role during telomere replication.
机译：复制端粒蛋白rTP是纤毛Euplotes crassus的核蛋白，似乎是一种新型的端粒复制因子。 rTP与两种蛋白具有广泛的氨基酸序列同一性，这两种蛋白结合并保护大核端粒免受纤毛虫Oxytricha和Euplotes的侵害。由于最广泛的保守区域位于端粒结合蛋白的DNA结合域之内，因此当首次鉴定出rTP时，它被认为是另一种结构性端粒结合蛋白。但是，随后的研究表明，rTP转录物仅在DNA复制过程中积累，并且rTP蛋白位于Euplotes大核内的DNA复制位点。我们现在已经在异源表达系统中表达了rTP，并检查了重组蛋白的DNA结合特性。我们显示，rTP特异地与Euplotes端粒DNA的G链结合，因此具有与Euplotes和Oxytricha端粒结合蛋白相同的DNA结合特征。但是，rTP绑定的其他方面是唯一的。特别地，该蛋白质表现出非常高的脱落率并且可以结合双链DNA以及端粒序列的内部片段。我们得出的结论是，rTP和端粒结合蛋白是一类蛋白质的成员，这些蛋白质具有保守的DNA结合基序，可以定制以结合端粒DNA的G链。但是，rTP独特的DNA结合特性表明该蛋白质已进化为在端粒复制过程中发挥特殊作用。

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