Identification and purification of a Drosophila protein that binds to the terminal 31-base-pair inverted repeats of the P transposable element.
Identification and purification of a Drosophila protein that binds to the terminal 31-base-pair inverted repeats of the P transposable element.
复制标题
鉴定和纯化与 P 转座元件末端 31 个碱基对反向重复序列结合的果蝇蛋白。
DOI：
10.1073/pnas.85.23.8929
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发表时间：
1988
影响因子：
11.1
通讯作者：
Rubin,GM
中科院分区：
文献类型：
--
作者：
Rio,DC;Rubin,GM
我们已经使用DNase I足迹法和部分分馏的果蝇KC组织培养细胞的核提取物，以确定与P转座因子的末端重复序列相互作用的DNA结合蛋白。我们已经确定了一种结合活性，它与直接邻近靶位点DNA复制的31个碱基对末端反向重复序列的区域特异性相互作用。结合发生在5'和3'反向末端重复序列两者上，而与复制的靶DNA的序列无关。UV光化学交联研究表明，结合活性存在于65-70 kDa的多肽中。生物化学分级分离和寡核苷酸亲和层析已被用来纯化的结合活性，以接近同质性，并确定在高度纯化的制剂中的66 kDa的多肽。发生结合的位点包含在P元件转位绝对需要的区域中，这表明该结合蛋白可能是参与P元件转位的细胞因子。
We have used DNase I footprinting and partially fractionated nuclear extracts from Drosophila Kc tissue culture cells to identify DNA-binding proteins that interact with the terminal repeats of P transposable elements. We have identified a binding activity that interacts specifically with a region of the 31-base-pair terminal inverted repeats that is directly adjacent to the duplication of target site DNA. Binding occurs to both the 5' and 3' inverted terminal repeats irrespective of the sequence of the duplicated target DNA. UV photochemical crosslinking studies suggest that the binding activity resides in a polypeptide of 65-70 kDa. Biochemical fractionation and oligonucleotide affinity chromatography have been used to purify the binding activity to near homogeneity and identify a polypeptide of 66 kDa in the highly purified preparations. The site to which binding occurs is included in a region absolutely required for P element transposition, suggesting that this binding protein may be a cellular factor involved in P element transposition.