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来自HeLa细胞的Alu家族重复序列结合蛋白,其与SV40病毒基因组的调控区域相互作用。

Alu-family repeat binding protein from HeLa cells which interacts with regulatory region of SV40 virus genome.

作者信息

Perelygina L M, Tomilin N V, Podgornaya O I

机构信息

Institute of Cytology, Academy of Sciences of the USSR, Leningrad.

出版信息

Mol Biol Rep. 1987;12(2):111-6. doi: 10.1007/BF00368878.

Abstract

Using a gel retardation assay the protein which binds selectively to the Alu-family repeat (AFR) has been identified and partially purified from HeLa cell nuclear extract. The protein (AFR-binding protein, ABP) forms multiple discrete complexes with AFR even in the presence of 200 to 2000-fold excess of non-specific (E. coli) DNA. The most stable complex has a relative mobility in 4% polyacrylamide gel (as compared to the free Alu-fragment) of 0.54. Heterogeneity of protein-DNA bands seen in the polyacrylamide gel suggests that ABP is able to form multimeric complexes with AFR. Competition experiments show that ABP does not interact with the RNA polymerase III promoter and with the TGGCA-sequence, but a high affinity binding site for ABP was found within a 660 bp restriction fragment containing the SV40 virus promoter and replication origin.

摘要

通过凝胶阻滞试验,已从HeLa细胞核提取物中鉴定并部分纯化了与Alu家族重复序列(AFR)选择性结合的蛋白质。即使存在200至2000倍过量的非特异性(大肠杆菌)DNA,该蛋白质(AFR结合蛋白,ABP)仍能与AFR形成多个离散的复合物。最稳定的复合物在4%聚丙烯酰胺凝胶中的相对迁移率(与游离Alu片段相比)为0.54。聚丙烯酰胺凝胶中观察到的蛋白质-DNA条带的异质性表明ABP能够与AFR形成多聚体复合物。竞争实验表明,ABP不与RNA聚合酶III启动子和TGGCA序列相互作用,但在一个包含SV40病毒启动子和复制起点的660 bp限制性片段内发现了ABP的一个高亲和力结合位点。

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