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首页> 外文期刊>Cell biology international. >COMBINED APPLICATION OF IN VIVO UV-CROSSLINKING AND IN VITRO LABEL TRANSFER IN THE EXAMINATION OF AU-RICH SEQUENCE BINDING PROTEIN RNA INTERACTIONS
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COMBINED APPLICATION OF IN VIVO UV-CROSSLINKING AND IN VITRO LABEL TRANSFER IN THE EXAMINATION OF AU-RICH SEQUENCE BINDING PROTEIN RNA INTERACTIONS

机译：体内UV交联和体外标签转移在检测Au-rich序列结合蛋白RNA相互作用中的组合应用

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摘要

A combination of in vivo UV light-induced crosslinking of nucleic acids to proteins and in vitro label transfer assay war applied to investigate specific interactions between AU-rich sequences (ARE) in the 3' UTR of lymphokine mRNAs and cytoplasmic AU-rich sequence element binding proteins (AUBP) in normal human lymphoblasts and MLA 144 gibbon lymphoid tumor cells. We demonstrate that a pool of cytoplasmic AUBP can be effectively crosslinked to RNA in vivo, suggesting a close association of these proteins with ARE sequences in the cytoplasm. We also show that the UV-crosslinked AUBP pool is markedly reduced in malignantly transformed MLA 144 cells compared with normal lymphoblasts, indicating weaker interaction between lymphokine ARE and AUBP in these tumor cells. Similar differences in AUBP-RNA associations were found between the membrane-bound polysomal subfractions of the two cell types where most of the AUBP activity was localized. We suggest that the decreased AUBP-mRNA association in MLA 144 cells might reflect a process concerned with disturbances of mRNA metabolism in the neoplastic phenotype.
机译：体内紫外光诱导的核酸与蛋白质的交联与体外标记物转移测定战争的结合，用于研究淋巴因子mRNA 3'UTR中富含AU的序列（ARE）与细胞质富含AU的序列元件之间的特异性相互作用正常人淋巴母细胞和MLA 144长臂猿淋巴瘤细胞中的结合蛋白（AUBP）。我们证明细胞质AUBP池可以在体内有效地交联到RNA，表明这些蛋白与细胞质中的ARE序列密切相关。我们还显示，与正常淋巴母细胞相比，在恶性转化的MLA 144细胞中，UV交联的AUBP池明显减少，表明淋巴因子ARE和AUBP在这些肿瘤细胞中的相互作用较弱。在大多数AUBP活性均位于其中的两种细胞类型的膜结合多体亚部分之间，发现了AUBP-RNA缔合的相似差异。我们建议，MLA 144细胞中AUBP-mRNA缔合减少可能反映了与肿瘤表型中mRNA代谢紊乱有关的过程。

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