Pecinka Ales, Kato Naohiro, Meister Armin, Probst Aline V, Schubert Ingo, Lam Eric
Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, 06466 Gatersleben, Germany.
J Cell Sci. 2005 Aug 15;118(Pt 16):3751-8. doi: 10.1242/jcs.02498. Epub 2005 Aug 2.
Fluorescent protein chromatin tagging as achieved by the lac operator/lac repressor system is useful to trace distinct chromatin domains in living eukaryotic nuclei. To interpret the data correctly, it is important to recognize influences of the tagging system on nuclear architecture of the host cells. Within an Arabidopsis line that carries lac operator/lac repressor/GFP transgenes, the transgene loci frequently associate with each other and with heterochromatic chromocenters. Accumulation of tagged fusion protein further enhances the association frequency. Independent experiments with a transgenic plant carrying another multi-copy transgene also revealed, independent of its transcriptional state, unusually high frequencies of association with each other and with heterochromatin. From these results we conclude that the lac operator/lac repressor chromatin tagging system may alter the spatial chromatin organization in the host nuclei (in particular when more than one insertion locus is present) and also that loci of homologous transgenic repeats associate more often with each other and with endogenous heterochromatin than normal euchromatic regions.
通过乳糖操纵子/乳糖阻遏物系统实现的荧光蛋白染色质标记,对于追踪活真核细胞核中不同的染色质结构域很有用。为了正确解读数据,认识标记系统对宿主细胞核结构的影响很重要。在携带乳糖操纵子/乳糖阻遏物/绿色荧光蛋白转基因的拟南芥品系中,转基因位点经常相互关联,并与异染色质着丝粒相关联。标记融合蛋白的积累进一步提高了关联频率。对携带另一个多拷贝转基因的转基因植物进行的独立实验也表明,与其转录状态无关,相互之间以及与异染色质的关联频率异常高。从这些结果我们得出结论,乳糖操纵子/乳糖阻遏物染色质标记系统可能会改变宿主细胞核中的空间染色质组织(特别是当存在多个插入位点时),并且同源转基因重复序列的位点相互之间以及与内源性异染色质的关联比正常常染色质区域更频繁。