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题目:: Maintenance of Long-Range DNA Interactions after Inhibition of Ongoing RNA Polymerase II Transcription
ID:
状态:: 发布时间May 26, 2010 , 更新时间 March 27, 2012 , 提交时间 Jan. 14, 2008,
物种:: Mus musculus
摘要:: A relationship exists between nuclear architecture and gene activity and it has been proposed that the activity of ongoing RNA polymerase II transcription determines genome organization in the mammalian cell nucleus. Recently developed 3C and 4C technology allowed us to test the importance of transcription for nuclear architecture. We demonstrate that upon transcription inhibition binding of RNA polymerase II to gene regulatory elements is severely reduced. However, contacts between regulatory DNA elements and genes in the β-globin locus are unaffected and the locus still interacts with the same genomic regions elsewhere on the chromosome. This is a general phenomenon since the great majority of intra- and interchromosomal interactions with the ubiquitously expressed Rad23a gene are also not affected. Our data demonstrate that without transcription the organization and modification of nucleosomes at active loci and the local binding of specific trans-acting factors is unaltered. We propose that these parameters, more than transcription or RNA polymerase II binding, determine the maintenance of long-range DNA interactions. Keywords: transcription inhibition 4C 4C experiments were performed in duplo, dye swap was included. Hybridisation was performed on dedicated micro-arrays. Probes (60-mers) were selected from the sequences 100 bp up –and downstream of HindIII sites. To prevent cross-hybridization, probes that had any similarity with highly abundant repeats (RepBase 10.09) were removed from the probeset. In addition, probes that gave more than two BLAST hits in the genome were also removed from the probeset. Sequence alignments were performed using MegaBLAST using the standard settings. A hit was defined as an alignment of 30 nt or longer.
实验种类:: other
样本量:: 10
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数据号:: E-GEOD-10170, GSE10170
数据状态:: 无法自动分析，您可以尝试手动分析数据。

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