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Transcription of adenovirus 2 major late and peptide IX genes under conditions of in vitro nucleosome assembly.

机译:在体外核小体组装条件下转录腺病毒2个主要晚期肽和IX肽基因。

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Plasmid DNA containing adenovirus 2 major late and peptide IX genes was assembled into nucleosomes in vitro, and the assembled nucleosomes were used as a template to study the regulatory mechanism of transcription initiation under these conditions. Neither the major late nor peptide IX genes was transcribed on the already-assembled nucleosomes. However, the major late gene, but not the peptide IX gene, was transcribed efficiently when the DNA was incubated with HeLa cell extracts prior to assembly into nucleosomes. These results indicate that prebinding of some component in the cell extracts to DNA is essential to activate transcription of the major late gene on nucleosomes assembled under the conditions used here. Since gene IX on the nucleosomes was not transcriptionally active regardless of preincubation of DNA with the extracts, some other component or another, different template structure which is not able to be identified in an in vitro system with deproteinized DNA template might be required for activation of peptide IX gene transcription. To know the function of the upstream sequences of the major late gene, effects of the deletion on transcription of nucleosomes were compared with that of deproteinized DNA. The result showed that depression of transcription by deleting the upstream sequences had more effect on nucleosomes than on deproteinized DNA.
机译:在体外将含有腺病毒2个主要晚期肽和IX肽基因的质粒DNA组装成核小体,并将组装的核小体用作模板来研究在这些条件下转录起始的调控机制。主要的晚期和肽IX基因都没有转录到已经组装好的核小体上。但是,当将DNA与HeLa细胞提取物温育后组装成核小体时,主要的晚期基因而不是肽IX基因被有效地转录。这些结果表明,细胞提取物中某些成分与DNA的预结合对于激活在此条件下组装的核小体上主要晚期基因的转录至关重要。由于核小体上的基因IX不具有转录活性,而与DNA与提取物的预温育无关,因此可能需要一些其他成分或其他不同的模板结构,这些模板结构无法在具有脱蛋白DNA模板的体外系统中鉴定出来。肽IX基因转录。为了了解主要晚期基因上游序列的功能,将缺失对核小体转录的影响与脱蛋白DNA的影响进行了比较。结果表明,通过删除上游序列来抑制转录对核小体的影响要大于对脱蛋白DNA的影响。

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