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一种具有末端特异性端粒蛋白DNA结合特性的非洲爪蟾卵因子。

A Xenopus egg factor with DNA-binding properties characteristic of terminus-specific telomeric proteins.

作者信息

Cardenas M E, Bianchi A, de Lange T

机构信息

Rockefeller University, New York, New York 10021-6399.

出版信息

Genes Dev. 1993 May;7(5):883-94. doi: 10.1101/gad.7.5.883.

Abstract

We have identified a Xenopus laevis protein factor that specifically recognizes vertebrate telomeric repeats at DNA ends. This factor, called Xenopus telomere end factor (XTEF), is detected predominantly in extracts of Xenopus eggs and ovaries, which are estimated to contain sufficient XTEF to bind approximately 3 x 10(7) DNA ends. In contrast, XTEF is much less abundant (approximately 90 per cell) in extracts of somatic cell nuclei. Mobility retardation analysis of the XTEF activity in egg extracts indicates that this factor binds the vertebrate telomeric repeat sequence (TTAGGG)2 when present in a single-stranded 3' overhang. Single-stranded 3' extensions of (TTTGGG)2, (AAAGGG)2, (TTACCC)2, or a nonrepetitive sequence fail to bind XTEF efficiently, whereas changes in the double-stranded sequence 5' to the TTAGGG repeat tail are tolerated. TTAGGG repeats are not recognized at internal position, at a 5' protruding end, or in double-stranded DNA. In addition, the factor does not bind RNA with single-stranded UUAGGG repeats at a 3' end. XTEF-DNA complexes form and are stable in high salt. The DNA-binding properties of XTEF resemble the characteristics of a class of terminus-specific telomere proteins identified previously in hypotrichous ciliates.

摘要

我们已经鉴定出一种非洲爪蟾蛋白质因子,它能特异性识别DNA末端的脊椎动物端粒重复序列。这种因子被称为非洲爪蟾端粒末端因子(XTEF),主要在非洲爪蟾卵和卵巢提取物中检测到,据估计这些提取物中含有的XTEF足以结合约3×10⁷个DNA末端。相比之下,XTEF在体细胞细胞核提取物中的丰度要低得多(每个细胞约90个)。对卵提取物中XTEF活性的迁移率阻滞分析表明,当脊椎动物端粒重复序列(TTAGGG)₂以单链3'突出端形式存在时,该因子能与之结合。(TTTGGG)₂、(AAAGGG)₂、(TTACCC)₂的单链3'延伸或非重复序列不能有效地结合XTEF,而TTAGGG重复序列尾部5'端双链序列的变化则是可以容忍的。TTAGGG重复序列在内部位置、5'突出端或双链DNA中均不被识别。此外,该因子不与3'端带有单链UUAGGG重复序列的RNA结合。XTEF - DNA复合物在高盐条件下形成且稳定。XTEF的DNA结合特性类似于先前在寡毛纲纤毛虫中鉴定出的一类末端特异性端粒蛋白的特征。

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