A genetic locus targeted to the nuclear periphery in living cells maintains its transcriptional competence.

A genetic locus targeted to the nuclear periphery in living cells maintains its transcriptional competence.
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DOI:
10.1083/jcb.200706060
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发表时间:
2008-01-14
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Spector DL
Spector DL
中科院分区:
其他
文献类型:
--
作者:
Kumaran RI;Spector DL

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外周核纤层主要但不完全与非活性染色质相关,被认为是核结构和基因表达的重要决定因素。我们在这里提出了一个诱导系统,以靶向遗传位点的核纤层在活的哺乳动物细胞。使用三维延时显微镜,我们确定靶向位点需要通过有丝分裂。一旦被靶向,该基因座在间期保持锚定到核外围,以及在通过随后的有丝分裂后保持锚定到子细胞中。在转录诱导后,基因表达机制的组成部分被招募到靶位点,我们在核周边看到了新生的转录本。在核纤层的转录诱导的动力学是类似于在内部核区域观察到的。这种新的细胞系统提供了一种强有力的方法来研究活细胞核周边基因功能的动态。
The peripheral nuclear lamina, which is largely but not entirely associated with inactive chromatin, is considered to be an important determinant of nuclear structure and gene expression. We present here an inducible system to target a genetic locus to the nuclear lamina in living mammalian cells. Using three-dimensional time-lapse microscopy, we determined that targeting of the locus requires passage through mitosis. Once targeted, the locus remains anchored to the nuclear periphery in interphase as well as in daughter cells after passage through a subsequent mitosis. Upon transcriptional induction, components of the gene expression machinery are recruited to the targeted locus, and we visualized nascent transcripts at the nuclear periphery. The kinetics of transcriptional induction at the nuclear lamina is similar to that observed at an internal nuclear region. This new cell system provides a powerful approach to study the dynamics of gene function at the nuclear periphery in living cells.
活细胞中 RNA 聚合酶 II 的转录周期。
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