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首页> 外文期刊>Molecular and Cellular Biology >PU.1 recruits a second nuclear factor to a site important for immunoglobulin kappa 3' enhancer activity.
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PU.1 recruits a second nuclear factor to a site important for immunoglobulin kappa 3' enhancer activity.

机译：PU.1将第二个核因子募集到对免疫球蛋白kappa 3'增强子活性重要的位点。

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摘要

PU.1 is a B-cell- and macrophage-specific transcription factor. By an electrophoretic mobility shift assay and dimethyl sulfate methylation interference assays, we show that PU.1 binds to DNA sequences within the immunoglobulin kappa 3' enhancer (kappa E3'). Binding of PU.1 to the kappa E3' enhancer assists the binding of a second tissue-restricted factor, NF-EM5, to an adjacent site. Binding of NF-EM5 to kappa E3' DNA sequences requires protein-protein interaction with PU.1 as well as specific protein-DNA interactions. This is the first known instance of PU.1 interacting with another cellular protein. NF-EM5 does not cofractionate with PU.1, suggesting that it is a distinct protein and is not a posttranslational modification of PU.1. UV-crosslinking studies and elution from sodium dodecyl sulfate-polyacrylamide gels indicate that NF-EM5 is a protein of approximately 46 kDa. Site-directed mutagenesis studies of the PU.1- and EM5-binding sites indicate that these sites play important roles in kappa E3' enhancer activity. By using a series of PU.1 deletion constructs, we have identified a region in PU.1 that is necessary for interaction with NF-EM5. This segment encompasses a 43-amino-acid region with PEST sequence homology, i.e., one that is rich in proline (P), glutamic acid (E), serine (S), and threonine (T).
机译：PU.1是B细胞和巨噬细胞特异性转录因子。通过电泳迁移率迁移测定和硫酸二甲酯甲基化干扰测定，我们显示PU.1结合至免疫球蛋白κ3'增强子（κE3'）内的DNA序列。 PU.1与Kappa E3'增强子的结合有助于第二组织限制因子NF-EM5与相邻位点的结合。 NF-EM5与Kappa E3'DNA序列的结合需要与PU.1的蛋白质-蛋白质相互作用以及特定的蛋白质-DNA相互作用。这是PU.1与另一种细胞蛋白相互作用的第一个已知实例。 NF-EM5与PU.1不共馏，表明它是一种独特的蛋白质，不是PU.1的翻译后修饰。紫外交联研究和十二烷基硫酸钠-聚丙烯酰胺凝胶洗脱表明，NF-EM5是大约46 kDa的蛋白质。对PU.1-和EM5结合位点的定点诱变研究表明，这些位点在Kappa E3'增强子活性中起重要作用。通过使用一系列的PU.1缺失构建体，我们已经确定了PU.1中与NF-EM5相互作用所必需的区域。该区段包含具有PEST序列同源性的43个氨基酸区域，即富含脯氨酸（P），谷氨酸（E），丝氨酸（S）和苏氨酸（T）的区域。

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