High resolution imaging reveals heterogeneity in chromatin states between cells that is not inherited through cell division.
High resolution imaging reveals heterogeneity in chromatin states between cells that is not inherited through cell division.
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DOI：
10.1186/s12860-016-0111-y
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发表时间：
2016-09-08
期刊：
影响因子：
--
通讯作者：
Owen-Hughes T
中科院分区：
文献类型：
--
作者：
Dickerson D;Gierliński M;Singh V;Kitamura E;Ball G;Tanaka TU;Owen-Hughes T
真核生物的基因组以染色质的形式存在，已知不同的染色质状态会影响基因的调控。染色质不是一种静态结构，但已知是动态的，在不同的细胞中是不同的。为了监测活细胞中染色质的组织结构，我们设计了能够识别特定操作符序列的荧光融合蛋白，以标记成对的共线基因座。然后使用荧光显微镜在三维上跟踪这些基因座的分离。我们建立了一个工作流程，用于测量两个荧光标记的共线基因座之间的距离，平均测量误差为63 nm。一般来说，观察到物理分离随着基因组分离的增加而增加。然而，染色质被压缩的程度在不同的基因组区域是不同的。没有观察到紧凑性与来自基因组数据集的染色质标记的分布或使用基于捕获的方法识别的接触者之间的相关性。细胞内和细胞间的空间分离也随着时间的推移而变化。单个基因座构象的差异可以在单个细胞中持续数分钟。在相关和不相关的子细胞对中，报告基因座的分离被发现是相似的。在活细胞中直接观察到的报告基因座的物理分离是高度动态的，无论是随着时间的推移，还是从细胞到细胞。然而，在一些染色体区域观察到了一致的分离差异，这些区域与已知的影响染色质状态的因素无关。我们的结论是，目前尚未确定的参数影响染色质的构型。我们还发现，尽管染色质状态的异质性可以在细胞间保持几分钟，但它不会通过细胞分裂而遗传。这可能导致了细胞间转录的异质性。本文的在线版本(doi：10.1186/s12860-0160111-y)包含补充材料，授权用户可以使用。
Genomes of eukaryotes exist as chromatin, and it is known that different chromatin states can influence gene regulation. Chromatin is not a static structure, but is known to be dynamic and vary between cells. In order to monitor the organisation of chromatin in live cells we have engineered fluorescent fusion proteins which recognize specific operator sequences to tag pairs of syntenic gene loci. The separation of these loci was then tracked in three dimensions over time using fluorescence microscopy. We established a work flow for measuring the distance between two fluorescently tagged, syntenic gene loci with a mean measurement error of 63 nm. In general, physical separation was observed to increase with increasing genomic separations. However, the extent to which chromatin is compressed varies for different genomic regions. No correlation was observed between compaction and the distribution of chromatin markers from genomic datasets or with contacts identified using capture based approaches. Variation in spatial separation was also observed within cells over time and between cells. Differences in the conformation of individual loci can persist for minutes in individual cells. Separation of reporter loci was found to be similar in related and unrelated daughter cell pairs. The directly observed physical separation of reporter loci in live cells is highly dynamic both over time and from cell to cell. However, consistent differences in separation are observed over some chromosomal regions that do not correlate with factors known to influence chromatin states. We conclude that as yet unidentified parameters influence chromatin configuration. We also find that while heterogeneity in chromatin states can be maintained for minutes between cells, it is not inherited through cell division. This may contribute to cell-to-cell transcriptional heterogeneity. The online version of this article (doi:10.1186/s12860-016-0111-y) contains supplementary material, which is available to authorized users.
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影响因子：
14.9
作者：
Baddeley D;Chagin VO;Schermelleh L;Martin S;Pombo A;Carlton PM;Gahl A;Domaing P;Birk U;Leonhardt H;Cremer C;Cardoso MC
通讯作者：
Cardoso MC
DOI：
10.1073/pnas.0810057105
发表时间：
2008-12-16
影响因子：
11.1
作者：
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10.5
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10.5
作者：
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影响因子：
10.6
作者：
Boulanger, Jerome;Kervrann, Charles;Salamero, Jean
通讯作者：
Salamero, Jean