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Analysis of multiple forms of nuclear factor I in human and murine cell lines.

机译:分析人类和鼠类细胞系中多种形式的核因子I。

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Nuclear factor I (NFI) is a group of related site-specific DNA-binding proteins that function in adenovirus DNA replication and cellular RNA metabolism. We have measured both the levels and forms of NFI that interact with a well-characterized 26-base-pair NFI-binding site. Five different NFI-DNA complexes were seen in HeLa nuclear extracts by using a gel mobility shift (GMS) assay. In addition, at least six forms of NFI were shown to cross-link directly to DNA by using a UV cross-linking assay. The distinct GMS complexes detected were composed of different subspecies of NFI polypeptides as assayed by UV cross-linking. Different murine cell lines possessed varying levels and forms of NFI binding activity, as judged by nitrocellulose filter binding and GMS assays. The growth state of NIH 3T3 cells affected both the types of NFI-DNA complexes seen in a GMS assay and the forms of the protein detected by UV cross-linking.
机译:核因子I(NFI)是一组相关的位点特异性DNA结合蛋白,在腺病毒DNA复制和细胞RNA代谢中起作用。我们已经测量了与特征明确的26个碱基对的NFI结合位点相互作用的NFI的水平和形式。通过使用凝胶迁移率变动(GMS)分析,在HeLa核提取物中发现了五种不同的NFI-DNA复合物。另外,通过使用紫外线交联测定法,显示出至少六种形式的NFI直接交联至DNA。检测到的独特的GMS复合物由NFI多肽的不同亚种组成,如通过UV交联测定的。如通过硝酸纤维素滤膜结合和GMS测定所判断的,不同的鼠细胞系具有不同水平和形式的NFI结合活性。 NIH 3T3细胞的生长状态会影响在GMS分析中发现的NFI-DNA复合物的类型以及通过UV交联检测到的蛋白质的形式。

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