Mapping interactions between mRNA export factors in living cells.

Mapping interactions between mRNA export factors in living cells.
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DOI:
10.1371/journal.pone.0067676
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Wilson SA
Wilson SA
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Teng IF;Wilson SA

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TREX复合物将核mRNA加工事件与随后输出到细胞质偶联。TREX还充当mRNA输出受体Nxf 1的结合平台。mRNA在细胞核内的转录和加工位点已被广泛研究。然而,关于TREX组装发生的位置以及Nxf 1在何处被募集到TREX以形成可输出的mRNP知之甚少。在这里,我们已经使用敏化发射Förster共振能量转移(FRET)和荧光寿命成像(FLIM)-FRET,在活细胞中产生的两个TREX亚基,Alyref和Chtop,与Nxf 1的相互作用的网站的空间图。在已知参与转录、剪接和外显子连接复合物形成的区域中的核斑点附近发现了输出因子的显著组装位点,这突出了mRNA输出与mRNP生物发生的密切耦合。
The TREX complex couples nuclear mRNA processing events with subsequent export to the cytoplasm. TREX also acts as a binding platform for the mRNA export receptor Nxf1. The sites of mRNA transcription and processing within the nucleus have been studied extensively. However, little is known about where TREX assembly takes place and where Nxf1 is recruited to TREX to form the export competent mRNP. Here we have used sensitized emission Förster resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM)-FRET, to produce a spatial map in living cells of the sites for the interaction of two TREX subunits, Alyref and Chtop, with Nxf1. Prominent assembly sites for export factors are found in the vicinity of nuclear speckles in regions known to be involved in transcription, splicing and exon junction complex formation highlighting the close coupling of mRNA export with mRNP biogenesis.
活细胞蛋白局部化的荧光共振能量转移(FRET)显微镜成像。
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