Transcribed DNA is preferentially located in the perichromatin region of mammalian cell nuclei
Transcribed DNA is preferentially located in the perichromatin region of mammalian cell nuclei
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DOI：
10.1016/j.yexcr.2010.10.026
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发表时间：
2011-02-15
影响因子：
3.7
通讯作者：
Fakan, Stanislav
中科院分区：
文献类型：
--
作者：
Niedojadlo, Janusz;Perret-Vivancos, Cecile;Fakan, Stanislav
转录 DNA 和产生的 RNA 的精确定位是细胞核功能结构的一个重要方面。为此，我们开发了一种与免疫电子显微镜相结合的新型原位杂交方法，使用源自总细胞或细胞质聚 (A+) RNA 的有义和反义 RNA 探针。这项新技术比使用 DNA 探针的经典原位杂交更加温和，并且能够很好地保存核结构。在固定和树脂包埋的 COS-7 细胞的超薄切片上进行，它以高分辨率揭示了编码细胞 mRNA 的基因的定位。定量分析表明，大多数转录的 DNA 集中在染色质周围区域，即染色体下致密染色质结构域和基本上没有 DNA 的染色质间空间之间的界面。产生的 RNA 主要存在于染色质周围区域和染色质间空间。这些结果表明，在哺乳动物细胞核中，染色质纤维被折叠，因此活性基因主要存在于染色质周围区域，这是最突出的转录位点。 (C) 2010 年，爱思唯尔公司出版。
The precise localization of transcribed DNA and resulting RNA is an important aspect of the functional architecture of the nucleus. To this end we have developed a novel in situ hybridization approach in combination with immunoelectron microscopy, using sense and anti-sense RNA probes that are derived from total cellular or cytoplasmic poly(A+) RNA. This new technology is much more gentle than classical in situ hybridization using DNA probes and shows excellent preservation of nuclear structure. Carried out on ultrathin sections of fixed and resin-embedded COS-7 cells, it revealed at high resolution the localization of the genes that code for the cellular mRNAs. Quantitative analysis shows that most transcribed DNA is concentrated in the perichromatin region, i.e. the interface between subchromosomal compact chromatin domains and the interchromatin space essentially devoid of DNA. The RNA that is produced is found mainly in the perichromatin region and the interchromatin space. These results imply that in the mammalian nucleus the chromatin fiber is folded so that active genes are predominantly present in the perichromatin region, which is the most prominent site of transcription. (C) 2010 Published by Elsevier Inc.